diff --git a/genesets/DEG_C2.CGP.M10970.txt b/genesets/DEG_C2.CGP.M10970.txt index cf19e5894e208ba53d22f4caff433554740f7836..2c11eacf33be93632fb1aaf802774540ff38c79e 100755 --- a/genesets/DEG_C2.CGP.M10970.txt +++ b/genesets/DEG_C2.CGP.M10970.txt @@ -1,5 +1,4 @@ CHUANG_OXIDATIVE_STRESS_RESPONSE_UP -> Genes up-regulated in MCF7 cells (breast cancer) after treatment with the oxydants: hydrogen peroxyde, menadione, and t-butyl hydroperoxyde [PubChem=784;4055;6410]. CTAGE5 CYP1B1 DDIT3 @@ -27,4 +26,4 @@ PSMD3 RCBTB1 RPL38 SLC35B3 -ZBTB4 \ No newline at end of file +ZBTB4 diff --git a/genesets/SupTab3_Consensus_Sigs.csv b/genesets/SupTab3_Consensus_Sigs.csv new file mode 100644 index 0000000000000000000000000000000000000000..4470e84bda5bc86370ace0c03784830b9731ebcb --- /dev/null +++ b/genesets/SupTab3_Consensus_Sigs.csv @@ -0,0 +1,288 @@ +Supplementary Table 3,,,,, +"Consensus marker genes for cell-type clusters from both initial 3' droplet-based and full-length plate-based scRNA-seq data (7,193 + 301 cells, in Sup. Tabs 1 and 2, respectively)",,,,, +"Thresholds: Fisher's combined FDR: 0.001 for plate-based and maximum FDR: 0.05 for droplet-based, Minimum log2 fold-change:0.25",,,,, +,,,,, +,,,,, +,,,,, +Basal,Club,Ciliated,Tuft,Neuroendocrine,Ionocyte +Aqp3,Scgb1a1,AU040972,Lrmp,Chga,Stap1 +Icam1,Bpifa1,Tmem212,Gnat3,Calca,Gm933 +Krt17,Scgb3a2,Dynlrb2,Gnb3,Cxcl13,Ascl3 +Krt5,Sftpd,Ccdc153,Plac8,Scg2,Foxi1 +Krt15,Alox15,Tppp3,Trpm5,Nov,Atp6v0d2 +Sfn,Pon1,1110017D15Rik,Gng13,Scg5,Moxd1 +Perp,Cyp4a12b,Fam183b,Ltc4s,Pcsk1,Gnas +Fxyd3,Wfdc1,Sec14l3,Rgs13,Dnajc12,Ldhb +Sdc1,Mgst1,Pltp,Hck,Uchl1,Rasd1 +Gstm2,Akr1c18,Rsph1,Alox5ap,Ddc,P2ry14 +F3,Sftpa1,Mlf1,Avil,Snca,Tfcp2l1 +Epas1,Lypd2,2410004P03Rik,Alox5,Snap25,Cd81 +Abi3bp,Ces1f,Tekt1,Ptpn6,Cib3,Cftr +Adh7,Gsto1,Cdh26,Atp2a3,Bex2,Slc12a2 +Id1,Tst,Cdhr3,Plk2,Guk1,Asgr1 +Upk1b,Ffar4,Nme5,Zfp428,Chgb,Parm1 +Hspa1a,Hp,Ccdc17,Ethe1,Ascl1,Atp6v1e1 +Dapl1,Chad,Adam8,Mctp1,Ngf,Iqgap1 +Hspb1,Selenbp1,Cyp2s1,Fxyd6,Cplx2,Rbpms +Zfp296,Mgat3,1700007K13Rik,1810046K07Rik,Igfbp5, +Dcn,Gabrp,Tm4sf1,Pik3r5,Pkib, +Rplp0,,Elof1,Cd300lf,Lrp11, +Fam107b,,Ly6a,Rassf6,Tcerg1l, +Tacstd2,,Sntn,Dclk1,Rbp4, +Rpl13,,Ppil6,Spib,Mthfd2, +Sat1,,Ifitm1,Pik3cg,Cd9, +Sult1d1,,Tctex1d4,Pde2a,Meis2, +Hpgd,,Aldh3b1,Matk,Ly6h, +S100a14,,Lrrc48,Vav1,Tmem158, +Rps18,,Foxj1,Sh2d7,Cacna2d1, +Emp1,,1700026L06Rik,Bmx,Syt7, +Sgk1,,Csrp2,Fyb,Ptp4a3, +Gsta4,,Pifo,Ccdc28b,, +Rps4x,,Acot1,Ptpn18,, +Rps5,,Dnali1,Bpgm,, +Oat,,Ccdc113,Itgb7,, +Rps8,,Pcp4l1,Ccdc109b,, +S100a16,,Spag6,Anxa4,, +Phlda3,,Capsl,Rgs2,, +Dusp1,,Lrrc51,Tspan6,, +Crlf1,,2610028H24Rik,Prss53,, +Sh3bgrl3,,Ldlrad1,Ly6g6f,, +Rps2,,Stmnd1,Inpp5d,, +Junb,,1700024G13Rik,Ivns1abp,, +Pcolce,,Dmkn,Nkd1,, +Cotl1,,Ccdc78,Sh3bgrl,, +Sord,,Chchd6,Pou2f3,, +Igfbp7,,1700016K19Rik,Sox9,, +Rpl8,,Calml4,Chn2,, +Rpl4,,Vpreb3,Krt18,, +,,Odf3b,Calm2,, +,,Ak7,Kctd12,, +,,1700003M02Rik,Etv1,, +,,1700026D08Rik,Coprs,, +,,Lrrc23,Cd47,, +,,BC051019,Espn,, +,,Tubb4b,Tmem245,, +,,Enkur,Pitpnc1,, +,,Bphl,Hap1,, +,,Spata18,Reep5,, +,,Fam216b,H2afj,, +,,Riiad1,Selm,, +,,1700088E04Rik,NA,, +,,1700007G11Rik,NA,, +,,Rsph4a,NA,, +,,Fam47e,NA,, +,,Agr3,NA,, +,,Fbxo36,NA,, +,,1700001C02Rik,NA,, +,,Cmbl,NA,, +,,Spag16,NA,, +,,Bok,NA,, +,,Ift43,NA,, +,,Ubxn10,NA,, +,,Tekt4,NA,, +,,Ccdc108,NA,, +,,Erich2,NA,, +,,6820408C15Rik,NA,, +,,Prdx5,NA,, +,,Fam92b,NA,, +,,Smim5,NA,, +,,Hdc,NA,, +,,Serpina9,NA,, +,,Ly6c1,NA,, +,,Aldh1a1,NA,, +,,Fhl1,NA,, +,,4833427G06Rik,NA,, +,,Rfk,NA,, +,,Dnaic2,NA,, +,,Ppp1r36,NA,, +,,Ak8,NA,, +,,B9d1,NA,, +,,Chchd10,NA,, +,,Zmynd10,NA,, +,,Rsph9,NA,, +,,Nek5,NA,, +,,Gm973,NA,, +,,Ccdc39,NA,, +,,Efhc1,NA,, +,,Hsph1,NA,, +,,Tuba1b,NA,, +,,Ubxn11,NA,, +,,Slc23a2,NA,, +,,Spa17,NA,, +,,Nat9,NA,, +,,Efcab10,NA,, +,,Cd177,NA,, +,,Hsp90aa1,NA,, +,,Iqca,NA,, +,,Dync2li1,NA,, +,,Ruvbl1,NA,, +,,Ppp1r32,NA,, +,,Spef1,NA,, +,,Ccdc81,NA,, +,,Dnajb13,NA,, +,,Acox2,NA,, +,,Fhad1,NA,, +,,Spag8,NA,, +,,Nudc,NA,, +,,Ccdc146,NA,, +,,Pih1d2,NA,, +,,Vwa3a,NA,, +,,Fam179a,NA,, +,,Fth1,NA,, +,,Ccdc151,NA,, +,,Lrrc36,NA,, +,,Cxcl17,NA,, +,,Kif9,NA,, +,,Ccdc37,NA,, +,,Mapk15,NA,, +,,Lhb,NA,, +,,Cd82,NA,, +,,Fam161a,NA,, +,,Rabl2,NA,, +,,Fank1,NA,, +,,Wdr63,NA,, +,,Glipr1,NA,, +,,Zfp474,NA,, +,,Lgals3bp,NA,, +,,Dpcd,NA,, +,,Ttc29,NA,, +,,Ttc16,NA,, +,,Mdh1b,NA,, +,,Ruvbl2,NA,, +,,Gipc2,NA,, +,,Arc,NA,, +,,Mak,NA,, +,,Dcdc2b,NA,, +,,Ccdc30,NA,, +,,1700028P14Rik,NA,, +,,Mycbp,NA,, +,,Oscp1,NA,, +,,Morn5,NA,, +,,Iqcg,NA,, +,,Eno4,NA,, +,,Ift74,NA,, +,,Lrrc34,NA,, +,,Cetn4,NA,, +,,Ankrd66,NA,, +,,Dnajc15,NA,, +,,Ulk4,NA,, +,,Tbata,NA,, +,,Traf3ip1,NA,, +,,Akr1b8,NA,, +,,6430531B16Rik,NA,, +,,Armc4,NA,, +,,Maats1,NA,, +,,Plekhb1,NA,, +,,Meig1,NA,, +,,Morn3,NA,, +,,Lrrc71,NA,, +,,1110004E09Rik,NA,, +,,Ift88,NA,, +,,Timp4,NA,, +,,Fam166b,NA,, +,,Ccdc121,NA,, +,,Usp18,NA,, +,,Ccdc33,NA,, +,,Slc23a1,NA,, +,,Gm166,NA,, +,,1810037I17Rik,NA,, +,,Syt5,NA,, +,,Dnaaf1,NA,, +,,Mns1,NA,, +,,Dusp14,NA,, +,,Wdr78,NA,, +,,Dynll1,NA,, +,,Gas8,NA,, +,,Ccdc114,NA,, +,,Crip2,NA,, +,,Ctxn1,NA,, +,,1700040L02Rik,NA,, +,,Tcp11,NA,, +,,Kif19a,NA,, +,,Ccdc65,NA,, +,,Tmem107,NA,, +,,Adgb,NA,, +,,Cib1,NA,, +,,Dnaic1,NA,, +,,Odc1,NA,, +,,Dnaaf3,NA,, +,,Cdkl4,NA,, +,,Mrps17,NA,, +,,Wdr34,NA,, +,,Slc9a3r1,NA,, +,,Kcnmb2,NA,, +,,Swi5,NA,, +,,Plin5,NA,, +,,Zc2hc1a,NA,, +,,Jam3,NA,, +,,Lrrc6,NA,, +,,Sptlc3,NA,, +,,Hydin,NA,, +,,Fam216a,NA,, +,,4932443I19Rik,NA,, +,,Ankrd42,NA,, +,,Wdr60,NA,, +,,Lrrc45,NA,, +,,Spef2,NA,, +,,Ezr,NA,, +,,Wnt11,NA,, +,,Fam213a,NA,, +,,Gtsf1l,NA,, +,,Lbp,NA,, +,,Wdr95,NA,, +,,Syne1,NA,, +,,Abhd3,NA,, +,,Tctex1d2,NA,, +,,Ttc39a,NA,, +,,Lrrc46,NA,, +,,Stk33,NA,, +,,Slc2a12,NA,, +,,Mipep,NA,, +,,Mst1,NA,, +,,Myl4,NA,, +,,Oaz2,NA,, +,,Ttc12,NA,, +,,Ccdc176,NA,, +,,Ift57,NA,, +,,Wdr54,NA,, +,,4930444P10Rik,NA,, +,,Tuba1a,NA,, +,,1700013F07Rik,NA,, +,,Mxra8,NA,, +,,D430036J16Rik,NA,, +,,Spag17,NA,, +,,Ddo,NA,, +,,Sri,NA,, +,,Ctsk,NA,, +,,Calm1,NA,, +,,4430402I18Rik,NA,, +,,2310007B03Rik,NA,, +,,Psmc3ip,NA,, +,,Nudt4,NA,, +,,Ttll9,NA,, +,,Dusp18,NA,, +,,Rfc2,NA,, +,,1700029J07Rik,NA,, +,,Naga,NA,, +,,Akap14,NA,, +,,Atp2a2,NA,, +,,Pttg1,NA,, +,,Casc1,NA,, +,,Ccdc60,NA,, +,,Dydc2,NA,, +,,Lrrc10b,NA,, +,,Mycbpap,NA,, +,,1700001L19Rik,NA,, +,,Dlec1,NA,, +,,Kif6,NA,, +,,Ropn1l,NA,, +,,Eml2,NA,, +,,Ech1,NA,, +,,Slc1a4,NA,, +,,Sftpc,NA,, +,,1700101E01Rik,NA,, +,,Efcab1,NA,, +,,Sirt3,NA,, +,,Pacrg,NA,, +,,Cdc14a,NA,, +,,Itln1,NA,, diff --git a/genesets/SupTab6_Krt13_Hillock.csv b/genesets/SupTab6_Krt13_Hillock.csv new file mode 100644 index 0000000000000000000000000000000000000000..1fa9fc8244896fd65a42b5a8ee323db9f1670e93 --- /dev/null +++ b/genesets/SupTab6_Krt13_Hillock.csv @@ -0,0 +1,5327 @@ +Supplementary Table 6,,,, +Related to Extended Data 4g: Krt4+/Krt13+ hillock-associated genes from 3' droplet-based scRNA-seq,,,, +Thresholds: all genes shown,,,, +,,,, +,,,, +,,,, +Gene,log2 fold-change (means),log2 fold-change (MAST),p,FDR +Ltf,4.371203488,1.997240249,2.12E-49,2.80E-47 +Krt4,2.883475949,2.715058937,3.95E-132,6.28E-129 +Upk3bl,2.789597952,2.775599468,1.09E-119,1.15E-116 +Nupr1,2.636378961,1.374198552,1.46E-41,1.45E-39 +S100g,2.625057175,2.520313317,3.50E-77,1.13E-74 +Lyz2,2.546423323,0.614993261,0.000231006,0.000456555 +Calml3,2.475765933,2.099366701,1.19E-75,3.60E-73 +Pglyrp1,2.434081355,1.60674033,7.79E-46,8.84E-44 +Crip1,2.388405052,2.069027685,6.52E-103,3.77E-100 +Ecm1,2.357414095,2.494416786,1.79E-98,8.76E-96 +S100a6,2.340319525,1.921351835,8.96E-152,2.85E-148 +Plac8,2.248865737,2.037148508,1.30E-67,3.06E-65 +Tppp3,2.23317182,2.510407089,5.57E-91,2.53E-88 +Cyp2a5,2.17096444,0.759483717,4.25E-12,5.37E-11 +Lgals3,2.077996831,2.427323601,1.72E-143,3.65E-140 +Cxcl17,2.038171513,1.030735714,3.53E-28,1.84E-26 +Ly6a,1.837604202,1.39606292,3.87E-51,5.48E-49 +Anxa1,1.835708894,2.195120887,1.87E-156,1.19E-152 +Ly6g6c,1.787286065,2.287473681,2.38E-55,3.78E-53 +Pdzk1ip1,1.750092129,1.376609155,8.05E-64,1.71E-61 +Cldn3,1.736300069,1.47012116,3.04E-106,1.93E-103 +2200002D01Rik,1.708508705,1.594216664,6.22E-79,2.20E-76 +Gsto1,1.66989588,1.366396368,2.10E-99,1.11E-96 +Sftpd,1.627169073,0.823349108,1.12E-21,3.95E-20 +Aldh1a1,1.615581755,0.659550713,1.32E-13,2.12E-12 +Pmm1,1.600405893,1.51168239,8.49E-85,3.38E-82 +Ndufa4,1.585913199,1.03861041,5.42E-60,1.04E-57 +Aqp5,1.567800927,1.816979092,9.19E-115,7.30E-112 +Slc16a11,1.563973486,0.704549261,2.00E-23,7.71E-22 +Scgb1a1,1.548737328,-1.054654362,2.05E-11,2.24E-10 +Chchd10,1.439711453,0.907958686,1.21E-23,4.75E-22 +Mal,1.432942435,1.438213019,5.51E-67,1.25E-64 +Upk1b,1.425739123,1.855816764,1.19E-63,2.44E-61 +Serpinb2,1.363477724,1.719984036,1.76E-47,2.20E-45 +Selenbp1,1.350855341,1.163362137,6.83E-55,1.06E-52 +Wfdc2,1.300652298,0.73103849,8.70E-27,4.04E-25 +Krt13,1.297049823,1.62029367,4.89E-33,3.31E-31 +Krt7,1.290614332,1.938782188,3.15E-122,4.00E-119 +Lrrc26,1.290586953,0.789865688,1.91E-35,1.48E-33 +Gabrp,1.290280684,0.77771119,2.85E-33,1.95E-31 +Cyp2b10,1.288995542,1.023290897,1.00E-40,9.65E-39 +Muc1,1.278783396,0.72805763,2.70E-27,1.28E-25 +Ly6e,1.265519729,0.942961381,6.55E-59,1.23E-56 +Pdlim1,1.225608541,1.260436821,2.07E-56,3.65E-54 +Krt15,1.222297944,1.779374151,2.59E-66,5.67E-64 +Ttc36,1.203599663,0.980892597,1.49E-40,1.38E-38 +Igfbp5,1.203541629,1.026713226,1.08E-38,9.31E-37 +S100a11,1.194691909,1.21670117,3.29E-115,2.99E-112 +Anxa3,1.173804934,1.371011691,3.54E-77,1.13E-74 +Ffar4,1.16253494,1.055926181,2.97E-51,4.29E-49 +Cd24a,1.152102004,0.882856505,7.53E-21,2.46E-19 +Qsox1,1.151083325,0.563475343,3.41E-14,5.80E-13 +Clca3b,1.131627199,0.879408754,2.68E-54,4.05E-52 +Alox15,1.11465741,0.597393309,1.61E-21,5.57E-20 +Socs2,1.10811153,0.967715386,4.58E-43,4.94E-41 +Txn1,1.100613032,0.818346722,2.52E-32,1.65E-30 +Tspo,1.091116077,1.128067989,1.03E-68,2.52E-66 +Porcn,1.074923076,1.0537818,1.79E-36,1.50E-34 +Cldn23,1.05859618,1.045825728,4.94E-40,4.49E-38 +Tacstd2,1.053937863,1.389993294,1.42E-55,2.31E-53 +Map1lc3a,1.030562527,1.01468104,2.79E-56,4.79E-54 +Ahnak,1.019957973,0.982600986,7.70E-46,8.84E-44 +Anxa2,1.005931457,1.558051857,1.45E-109,1.03E-106 +Krt23,0.99687152,0.847909907,1.23E-28,6.68E-27 +1810037I17Rik,0.974937316,0.894585759,9.64E-36,7.57E-34 +Ces1f,0.969763476,0.345538642,7.68E-15,1.40E-13 +S100a10,0.954702046,1.770067525,4.71E-61,9.36E-59 +Muc4,0.953978097,0.812118201,5.19E-30,3.03E-28 +Mgst3,0.945952909,1.133024383,6.22E-44,6.82E-42 +Lypd2,0.937773557,0.337824172,7.49E-06,2.18E-05 +Tspan1,0.929865493,0.442746632,1.20E-08,6.90E-08 +Gsta4,0.929786044,1.190897298,1.65E-83,6.16E-81 +Cyba,0.91956081,0.620934448,2.02E-27,9.71E-26 +St3gal4,0.918893788,0.901165362,1.83E-42,1.94E-40 +Ctsh,0.910706194,0.777605097,2.34E-35,1.79E-33 +Cldn7,0.90457056,0.978312028,2.84E-50,3.85E-48 +Ypel3,0.900066516,0.789060168,2.17E-34,1.59E-32 +Adh7,0.898427476,1.128975278,7.58E-51,1.05E-48 +Tspan8,0.897599234,0.901707015,5.95E-28,3.05E-26 +Fmo3,0.896108264,0.423253268,2.00E-12,2.68E-11 +Lmo7,0.892474596,0.814248664,7.63E-26,3.35E-24 +Sub1,0.889348458,0.720605451,3.59E-31,2.26E-29 +Capg,0.879521526,1.096395228,1.48E-45,1.65E-43 +Cd44,0.870810067,0.882754332,1.68E-33,1.20E-31 +Ebp,0.869112144,0.802133586,2.88E-30,1.73E-28 +Alcam,0.86827811,0.632843245,2.41E-28,1.28E-26 +S100a14,0.866587556,1.112455256,1.36E-53,2.02E-51 +Taldo1,0.863789861,0.77983576,2.20E-39,1.97E-37 +Ifitm1,0.849831918,1.09227457,9.17E-36,7.29E-34 +Crip2,0.843335977,1.074021961,2.90E-48,3.76E-46 +Trp53inp2,0.84104974,0.789034015,2.17E-28,1.17E-26 +Cwh43,0.840379884,0.95214878,3.24E-38,2.75E-36 +Cmpk1,0.827305108,0.731710963,3.41E-27,1.61E-25 +Neat1,0.818735448,0.551991992,2.14E-15,4.20E-14 +Tmsb4x,0.807745933,1.139481666,3.44E-56,5.75E-54 +Ces1d,0.804905003,0.898131643,3.66E-30,2.16E-28 +Krt19,0.794526333,1.271993149,1.55E-89,6.59E-87 +Clic1,0.793190583,0.87935758,1.26E-46,1.51E-44 +Polr2f,0.789769521,0.577216203,2.33E-19,6.91E-18 +Clic3,0.784108248,0.787543894,6.68E-28,3.40E-26 +Mrpl14,0.769488254,0.519020512,4.77E-16,1.02E-14 +Myh14,0.768228861,0.489674501,3.07E-15,5.88E-14 +S100a1,0.76199469,0.493993847,3.70E-12,4.72E-11 +Ly6d,0.752698241,0.788240171,1.00E-15,2.03E-14 +Fth1,0.745891053,0.741920067,6.47E-46,7.62E-44 +Krt8,0.740280784,1.169984136,2.68E-69,6.82E-67 +Tst,0.738248347,0.186165823,0.002156685,0.003346308 +Spint2,0.737294865,0.690485847,1.21E-46,1.48E-44 +Kctd14,0.735383573,0.662637189,5.74E-29,3.20E-27 +Rab25,0.714352965,0.76710415,1.96E-34,1.45E-32 +Prdx5,0.711861156,0.621110872,2.81E-16,6.10E-15 +Prr13,0.707096455,0.694790969,3.55E-32,2.28E-30 +Cyb5a,0.705088238,0.604360207,3.85E-32,2.45E-30 +Pllp,0.703824059,0.692012757,1.60E-29,9.11E-28 +Agr2,0.703116433,0.273413409,0.031796216,0.038889217 +Gprc5a,0.69781096,1.015837642,1.05E-47,1.34E-45 +Emp2,0.697236819,0.746551681,9.06E-23,3.31E-21 +AU040972,0.694985987,0.730521948,6.60E-20,2.00E-18 +Cmas,0.685811921,0.574685305,6.82E-24,2.71E-22 +Pmp22,0.685275009,0.9738596,1.16E-32,7.68E-31 +Pdlim2,0.684833643,0.669799346,1.13E-29,6.51E-28 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+Uxs1,0.019792905,0.025643227,0.002124862,0.003301765 +Lage3,0.01975001,0.058506789,3.33E-08,1.70E-07 +Ssrp1,0.019693225,0.101387239,8.99E-09,5.32E-08 +Sf3b3,0.019666638,0.005783621,0.032491938,0.039686715 +Mdp1,0.01965173,0.061609679,1.02E-05,2.86E-05 +Nupr1l,0.019627799,-0.041141678,0.00082896,0.001420308 +Ash1l,0.019557582,-0.004296221,0.000106635,0.000228581 +Tbc1d20,0.019549067,-0.00391289,5.74E-06,1.72E-05 +Fam220a,0.019540918,0.121166766,0.00190362,0.002987175 +Pdcd2,0.019525974,-0.003247026,4.29E-05,0.000101996 +Man1b1,0.019497327,0.026878143,8.64E-06,2.47E-05 +Eif6,0.019482153,0.309803396,1.61E-08,8.96E-08 +Rps6kb2,0.019431946,0.04660012,0.000291711,0.000560847 +Gtf2f2,0.01943045,0.055635847,0.003585507,0.00528478 +Anapc11,0.019369333,0.165689181,0.0003471,0.000655866 +Mcm6,0.019245165,0.0155787,0.031110779,0.038102168 +Ywhag,0.019144847,0.073769376,0.020574727,0.025989129 +Rars,0.019120357,0.077570072,0.000526741,0.000948493 +Snap23,0.019062953,0.073955371,0.012438721,0.016423139 +Abhd11,0.019050612,0.137747536,2.79E-10,2.34E-09 +Izumo4,0.019037112,0.013357413,7.44E-05,0.000165978 +Tollip,0.019013576,-0.00395257,2.54E-06,8.32E-06 +Med9,0.018928486,0.00290699,0.001157723,0.00191648 +Rps28,0.018866181,0.079103645,1.14E-06,4.06E-06 +Cdc42se2,0.018861863,0.030196694,0.026959687,0.033358679 +Naa20,0.018803183,0.145293365,0.001288553,0.002112708 +Nsmce2,0.018729138,0.088143186,2.11E-05,5.44E-05 +Tbcd,0.018721196,0.020257613,0.001306939,0.002139414 +Itln1,0.018608028,-0.051498539,2.06E-05,5.35E-05 +Rtp4,0.018399678,-0.048830228,0.000123731,0.000260918 +Fasn,0.018341828,0.010515474,0.016007766,0.020659374 +Snapin,0.018298037,0.103257571,0.000302188,0.000579239 +Arhgef19,0.0182654,0.015641092,0.015659477,0.020275707 +Med17,0.018218563,0.011872703,0.000377111,0.000706459 +Seh1l,0.018208235,0.078726144,0.01390681,0.018180331 +Ufsp1,0.018192526,0.002944866,0.00202929,0.003165633 +Prkag1,0.018138552,0.098297554,0.000160478,0.000329451 +Pik3r4,0.01813357,0.008410783,0.000812023,0.001393542 +Gorasp2,0.01812236,0.036976622,0.000718272,0.001247841 +Iscu,0.018068399,0.121523385,1.53E-05,4.11E-05 +Rbbp9,0.018061244,0.007985914,0.001818876,0.002870483 +Wbscr22,0.018027083,0.059079332,0.000209132,0.000418001 +Noc2l,0.017999275,0.042039855,0.003935046,0.005740802 +Serpinf1,0.017955909,-0.179270194,7.12E-09,4.29E-08 +Gpr107,0.017937296,-0.031703406,0.000157571,0.000324321 +Mgrn1,0.017826298,-0.043895711,0.011501181,0.015270879 +Bpgm,0.017771252,0.126288734,0.008041569,0.010998791 +Imp4,0.01770657,0.071613048,2.91E-06,9.36E-06 +Lims1,0.017685814,0.064463662,0.001490436,0.002404207 +Tmem19,0.017677928,0.052676535,8.55E-05,0.00018809 +Plscr1,0.0176335,0.251183201,0.000121116,0.000256083 +Stx16,0.017539719,0.028525812,0.023339302,0.029197081 +Ino80b,0.017537716,0.040845307,0.00016987,0.000346942 +Ikbip,0.017526142,-0.0106363,2.36E-05,6.01E-05 +Cbx1,0.017525168,0.031900967,0.004482595,0.006462492 +Cmtm6,0.017341221,-0.024714933,2.97E-07,1.21E-06 +Rpl22,0.017330983,-0.035219736,9.19E-05,0.000200452 +Vangl1,0.017185159,0.012013658,0.022047521,0.027728343 +Mef2a,0.017157783,0.060055179,0.010638604,0.01421162 +Herc1,0.017121001,0.003859709,0.034191101,0.041538758 +Pum1,0.017117058,0.023144239,0.00093506,0.001579544 +Kars,0.017047532,-0.008914141,0.00035466,0.000668338 +Cs,0.016939563,0.112077899,0.005506473,0.007784212 +Ube2w,0.016929012,0.025611933,0.000838479,0.001434685 +Znhit1,0.016915801,0.005038305,1.58E-08,8.85E-08 +Cog2,0.016912182,0.011971862,0.011401751,0.015148347 +Csrnp1,0.016902832,0.14737307,0.003207769,0.00477226 +Rbp4,0.016760505,-0.142933218,1.94E-11,2.14E-10 +Polr2a,0.016559548,0.054622912,0.00196006,0.003065154 +Tiam1,0.016485942,0.025818322,0.040261077,0.048304178 +Trpc4ap,0.016484601,-0.036179533,5.19E-13,7.75E-12 +Trmt6,0.016482736,0.008854228,0.012360177,0.016326215 +Brwd1,0.016479738,0.007511944,1.47E-05,3.97E-05 +Tcta,0.016398837,0.015726262,5.80E-06,1.74E-05 +Wdr83,0.016372273,0.036378016,0.000408305,0.000757754 +Ilf2,0.016260515,0.078657679,0.000104086,0.000223644 +Mrps22,0.016200704,0.022921412,0.035993141,0.043553344 +Rbck1,0.016151674,0.08734939,0.01363805,0.017862026 +Lemd1,0.016149057,0.049377212,0.036368624,0.04397423 +Paip2,0.016118668,0.306688241,6.33E-09,3.84E-08 +Tmem184a,0.016086799,0.018600043,6.85E-05,0.000154506 +Znhit2,0.016086658,0.040382558,0.005174783,0.007369373 +Bhlhe40,0.016060629,0.380939494,5.28E-09,3.25E-08 +Il17rc,0.01604583,0.036290924,6.78E-08,3.20E-07 +Whsc1l1,0.015933403,0.021753336,1.41E-08,7.97E-08 +Rcn2,0.015915642,0.053824081,0.000769277,0.001325549 +Oxld1,0.015909298,-0.053630469,7.47E-05,0.000166692 +Bckdha,0.015891634,0.05219855,1.52E-07,6.64E-07 +Syngr1,0.015754694,-0.071557325,4.83E-12,6.01E-11 +Atp2c2,0.015747014,-0.033639524,3.18E-07,1.29E-06 +Adipor2,0.015710501,-0.061945008,4.45E-06,1.36E-05 +Ccnl2,0.015709382,0.046296167,0.00107188,0.00178553 +Nfib,0.015656594,-0.025899574,1.10E-07,4.95E-07 +Commd10,0.01565079,0.095639965,0.006407256,0.008936987 +Tmem251,0.01563841,0.04525447,0.015872324,0.020513655 +Gtpbp8,0.015604973,-0.021323776,0.000757402,0.001308632 +Shkbp1,0.015580816,0.036207537,0.0001144,0.000243257 +Sarnp,0.015552129,0.197747424,6.86E-09,4.15E-08 +Serpinb12,0.01550104,0.007641917,0.014339031,0.018709921 +Prom1,0.015469625,-0.126310385,8.06E-09,4.81E-08 +Wdr6,0.015467217,0.006409266,0.024248132,0.030240225 +Tmem33,0.015439989,0.135140718,2.73E-05,6.86E-05 +Asb3,0.015413677,0.023509393,0.000190989,0.000385194 +Ptgr2,0.015349922,-0.040576206,1.72E-05,4.54E-05 +Itga6,0.015315098,0.165882157,0.006046242,0.008483146 +Xab2,0.015273982,-0.002974239,0.000180544,0.000365781 +Ensa,0.015257969,0.024363069,0.003437848,0.005086 +Uchl5,0.015164662,0.041492317,0.000493058,0.000893659 +1600012H06Rik,0.015152158,0.008874308,0.013346275,0.017515953 +Zfp652,0.014973899,-0.070212418,2.21E-05,5.68E-05 +Flywch2,0.014943653,-0.047766666,0.00010508,0.000225579 +Lyrm5,0.014939748,0.010297342,0.001286996,0.002111244 +Alkbh6,0.014937161,-0.052523161,0.00508603,0.007249474 +Man2c1,0.014920641,0.03791534,0.013561356,0.017768896 +Ndufb6,0.014905448,0.102374793,1.24E-05,3.42E-05 +Plrg1,0.014853243,0.044779702,0.017759016,0.022730397 +Cdkn2c,0.014846164,-0.021263368,0.002978825,0.004471401 +Igsf5,0.014825637,-0.000594872,0.002574138,0.003911018 +Tspyl1,0.014823803,0.014973477,0.017941746,0.022931975 +Mrpl13,0.014818039,-0.009852759,8.44E-08,3.88E-07 +Ptrh2,0.014809801,0.075469069,0.011611488,0.015401264 +Zcchc11,0.014795087,0.008495246,0.033136101,0.04041918 +Rbm18,0.014794925,0.068460508,0.00035389,0.000667281 +Fam96b,0.01477844,0.143148665,3.11E-05,7.67E-05 +Ikbkg,0.014776735,-0.023142753,0.000292635,0.000562284 +Usmg5,0.014746816,0.07194583,6.64E-05,0.000150412 +Cdc42ep4,0.014740424,0.091452155,0.000317447,0.000605568 +Rheb,0.014712737,0.214547733,3.95E-13,6.03E-12 +Nav2,0.014605645,-0.027593601,7.73E-05,0.000171739 +Ube2e3,0.014549846,0.058377703,1.93E-05,5.03E-05 +Dhps,0.014548489,-0.028843317,0.000362875,0.0006822 +Fam162a,0.014516214,0.020697568,4.28E-11,4.40E-10 +Arfgap2,0.014487957,0.007272193,4.80E-06,1.46E-05 +Scrib,0.014272584,0.02357608,2.75E-07,1.13E-06 +Pnisr,0.014041576,-0.012697763,0.016201159,0.020904721 +Chd4,0.014010706,0.027204219,0.00010858,0.000232279 +Insig1,0.013907432,-0.009528147,0.000228739,0.00045278 +Ubl7,0.013871664,0.068434688,9.57E-05,0.000207828 +Med10,0.013764051,0.050496256,2.90E-05,7.22E-05 +Exosc5,0.013758785,0.092018741,7.16E-05,0.00016084 +Zzef1,0.013665711,0.029880921,0.022978356,0.028790848 +Decr1,0.013664235,-0.000460548,4.15E-07,1.64E-06 +Zc2hc1a,0.013651686,0.116224014,0.004477137,0.006458288 +Acox1,0.013611497,0.031066082,3.02E-06,9.65E-06 +Baiap2l2,0.013600417,0.052544157,0.000954259,0.001607705 +2010107G23Rik,0.013540173,0.1013274,8.16E-05,0.00018031 +Ncln,0.013523899,0.026430184,0.003949484,0.005759852 +Mrpl2,0.013472871,0.000445093,1.65E-10,1.47E-09 +Sike1,0.013465567,0.037691753,4.67E-05,0.000109886 +Limk2,0.013400558,-0.018856263,6.14E-08,2.93E-07 +Slc44a2,0.013282797,0.091163481,0.005247281,0.007460923 +Ccng1,0.013262072,0.006187261,6.99E-08,3.28E-07 +Mul1,0.013257517,0.004034056,0.006855185,0.009500757 +Mat2b,0.013176257,0.075553924,1.73E-07,7.50E-07 +Gjb4,0.013118505,0.147711179,0.002651558,0.004020003 +Gpr87,0.012997793,0.171940493,0.00116606,0.001929277 +Uri1,0.012992149,0.026372066,0.005010052,0.007146114 +Tsnax,0.01291019,0.114490254,1.60E-07,6.94E-07 +Thap11,0.012886833,0.0312262,0.000405325,0.000753102 +Ndufaf4,0.012883473,0.014748854,0.000660107,0.001161671 +Gpr137,0.01285576,0.03902306,0.000234111,0.000461688 +Ctnnbl1,0.012842205,0.03226467,1.89E-05,4.96E-05 +Gmnn,0.012774556,-0.319710207,5.51E-15,1.02E-13 +Morf4l2,0.012710646,0.142720927,2.50E-05,6.33E-05 +Slc46a3,0.01258668,-0.026996638,6.97E-05,0.000156844 +Sf3a1,0.012583149,0.004863433,0.000504008,0.000910909 +Ndufb11,0.012555988,-0.012153434,0.000211222,0.000421648 +Nae1,0.012519309,-0.001415208,0.01009179,0.013560909 +Eif5,0.012473721,0.160500194,1.15E-05,3.18E-05 +Cxxc1,0.012426216,0.051009158,0.012271228,0.01621883 +Synpo,0.012415419,0.001257678,1.54E-05,4.14E-05 +Sdha,0.012413058,-0.012320446,6.31E-07,2.40E-06 +Dlat,0.012364428,0.084875587,0.018450093,0.023506129 +Polr1d,0.012330219,0.156339289,7.43E-08,3.46E-07 +Rpa3,0.01232436,-0.026483996,3.02E-07,1.23E-06 +Nkiras2,0.01226806,0.144719248,0.003523043,0.005198737 +Swi5,0.012261985,0.076246086,0.000388966,0.00072664 +Cpne8,0.012253356,0.051511103,0.005568584,0.007863276 +Ube2h,0.012172536,-0.008934639,7.23E-05,0.000162081 +Lypd6b,0.012132584,0.10913414,0.001728779,0.002739855 +Syncrip,0.012036489,-0.032808834,0.001380362,0.002245868 +Psmc2,0.01197174,0.098105864,3.84E-08,1.94E-07 +Rmnd5b,0.011925027,0.034369735,0.004988558,0.007123312 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+Safb,0.011230629,0.053467085,0.004893385,0.006997 +Fundc2,0.011208294,-0.004662674,8.07E-06,2.33E-05 +Zfand5,0.011045773,0.575982864,1.07E-10,1.01E-09 +Nrarp,0.011033708,0.104647964,0.001049962,0.001753855 +Asgr1,0.010970247,-0.167443626,2.74E-10,2.31E-09 +Tmem55a,0.010967477,0.022285127,0.003180941,0.004738999 +Tmem246,0.010965293,-0.01691493,0.000270626,0.00052459 +Eftud2,0.010944718,0.012780997,0.010800413,0.014406592 +Rpp14,0.010904012,0.027072645,1.30E-05,3.53E-05 +Cog8,0.010874657,0.035392865,6.21E-06,1.85E-05 +Tssc1,0.010799201,0.047156095,0.003346378,0.004964423 +Sergef,0.010700992,-0.038585079,0.010380391,0.013892974 +Gtpbp4,0.01069764,0.038420465,5.08E-05,0.000118247 +Lrrc8d,0.010673525,0.034503801,0.031275375,0.038274271 +Pqlc3,0.010534201,0.044121433,0.000488392,0.000886212 +Cd151,0.01051053,0.384694148,5.10E-09,3.15E-08 +Exoc3l4,0.01049898,-0.040358042,5.01E-06,1.52E-05 +Stard7,0.010457244,0.011640077,0.000128536,0.000270066 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+Gstm2,-1.027660036,-0.441568823,1.74E-05,4.60E-05 +Dcn,-1.033450621,-0.241015395,5.83E-18,1.53E-16 +Scgb3a2,-1.065078259,-1.788156064,2.57E-36,2.10E-34 +Rps4x,-1.070883306,-0.301305387,0.001708078,0.002710661 +Socs3,-1.109837142,-0.296756409,4.64E-09,2.89E-08 +Eef1g,-1.149710247,-0.437972402,9.71E-11,9.26E-10 +Crlf1,-1.156910774,-0.291048337,7.78E-08,3.61E-07 +Csrp1,-1.168228271,0.198589711,0.006101555,0.008555091 +Icam1,-1.177473548,-0.28246424,5.65E-06,1.70E-05 +Timp3,-1.183800689,-0.290056807,2.05E-16,4.52E-15 +Mt1,-1.189611344,-0.584942052,1.93E-18,5.32E-17 +Apoe,-1.225673839,-0.309081265,8.21E-09,4.89E-08 +Sfn,-1.248338935,0.762522458,2.83E-09,1.88E-08 +Sult1d1,-1.255032957,-1.249895909,2.70E-27,1.28E-25 +Rpl13,-1.28888753,-0.148967986,0.013211295,0.017349543 +Dnajb1,-1.323235005,0.403906848,2.74E-05,6.88E-05 +Igfbp7,-1.422419761,-0.497129455,3.62E-09,2.31E-08 +Atf3,-1.555657768,0.363769954,0.002234847,0.003452745 +Dapl1,-1.580876081,-0.243762125,0.035875874,0.043444509 +Scgb3a1,-1.581521583,-2.672410666,7.77E-75,2.15E-72 +Krt5,-1.624705022,-0.345614662,0.000511577,0.000923277 +Gadd45g,-1.766051524,-0.414367186,1.09E-11,1.26E-10 +Hspa8,-1.842390042,0.008383574,0.000331265,0.000629096 +Cyr61,-1.93928028,-0.2652897,0.006894105,0.009550535 +Hspa1a,-1.977226186,0.311311038,1.11E-13,1.78E-12 +Bpifa1,-2.098478366,-2.837998161,1.60E-75,4.62E-73 +Reg3g,-2.463513883,-2.861396445,1.54E-74,4.09E-72 diff --git a/r.scripts/sc-TissueMapper.R b/r.scripts/sc-TissueMapper.R index db19abd99e8bccf2b00fc2a888d773b5a4f022e4..37ad9f9655fe9d440bc8cb3ea3718be8e3c98f4b 100644 --- a/r.scripts/sc-TissueMapper.R +++ b/r.scripts/sc-TissueMapper.R @@ -491,14 +491,27 @@ scQuSAGE <- function(sc10x,gs,res.use=0.1,ds=25000,nm="Lin",folder="lin"){ number.clusters <- length(unique(sc10x@ident)) labels <- paste0("Cluster_",as.vector(factor(sc10x@ident))) - if ((ncol(sc10x@data)>ds & ds!=0)){ - rnd <- sample(1:ncol(sc10x@data),ds) - data <- sc10x@data[,rnd] - labels <- labels[rnd] - } else { - data <- sc10x@data + + #if ((ncol(sc10x@data)>ds & ds!=0)){ + # rnd <- sample(1:ncol(sc10x@data),ds) + # data <- sc10x@data[,rnd] + # labels <- labels[rnd] + #} else { + # data <- sc10x@data + #} + + cell.sample <- NULL + for (i in 1:number.clusters){ + cell <- names(sc10x@ident[sc10x@ident==i]) + if (length(cell)>ds & ds!=0){ + rnd <- sample(1:length(cell),ds) + cell <- cell[rnd] + } + cell.sample <- c(cell.sample,cell) } + data <- sc10x@data[,colnames(sc10x@data) %in% cell.sample] data <- as.data.frame(as.matrix(data)) + labels <- labels[colnames(sc10x@data) %in% cell.sample] #Make labels for QuSAGE clust <- list() @@ -541,7 +554,7 @@ scQuSAGE <- function(sc10x,gs,res.use=0.1,ds=25000,nm="Lin",folder="lin"){ if (max(results.cor[results.cor[,4]==i,][,2])>=0){ results.clust.id <- rbind(results.clust.id,results.cor[results.cor[,4]==i,][which.max(results.cor[results.cor[,4]==i,][,2]),]) } else { - results.clust.id <- rbind(results.clust.id,c("Unknown",0,0,i)) + results.clust.id <- rbind(results.clust.id,data.frame(pathway.name="Unknown",log.fold.change=0,FDR=0,Cluster=i)) } } rownames(results.clust.id) <- NULL @@ -601,7 +614,7 @@ scQuSAGE <- function(sc10x,gs,res.use=0.1,ds=25000,nm="Lin",folder="lin"){ merge.cluster <- c(merge.cluster,"Unknown") }} sc10x@ident <- plyr::mapvalues(x=sc10x@ident,from=1:number.clusters,to=merge.cluster) - sc10x@ident <- factor(sc10x@ident,levels=names(gs)) + sc10x@ident <- factor(sc10x@ident,levels=c(names(gs),"Unknown")) sc10x <- StashIdent(object=sc10x,save.name=nm) postscript(paste0("./analysis/tSNE/",folder,"/tSNE_",nm,".eps")) @@ -638,14 +651,29 @@ scQuSAGEsm <- function(sc10x,gs,ds=25000,nm="Lin",folder="lin"){ clusters <- unique(sc10x@ident) labels <- as.vector(factor(sc10x@ident)) - if ((ncol(sc10x@data)>ds & ds!=0)){ - rnd <- sample(1:ncol(sc10x@data),ds) - data <- sc10x@data[,rnd] - labels <- labels[rnd] - } else { - data <- sc10x@data + + #if ((ncol(sc10x@data)>ds & ds!=0)){ + # rnd <- sample(1:ncol(sc10x@data),ds) + # data <- sc10x@data[,rnd] + # labels <- labels[rnd] + #} else { + # data <- sc10x@data + #} + #data <- as.data.frame(as.matrix(data)) + + cell.sample <- NULL + for (i in clusters){ + cell <- names(sc10x@ident[sc10x@ident==i]) + if (length(cell)>ds & ds!=0){ + rnd <- sample(1:length(cell),ds) + cell <- cell[rnd] + } + cell.sample <- c(cell.sample,cell) } + data <- sc10x@data[,colnames(sc10x@data) %in% cell.sample] data <- as.data.frame(as.matrix(data)) + labels <- labels[colnames(sc10x@data) %in% cell.sample] + #Make labels for QuSAGE clust <- list() @@ -744,14 +772,28 @@ scQuSAGElg <- function(sc10x,gs,ds=25000,nm="Lin",folder="lin"){ clusters <- unique(sc10x@ident) labels <- as.vector(factor(sc10x@ident)) - if ((ncol(sc10x@data)>ds & ds!=0)){ - rnd <- sample(1:ncol(sc10x@data),ds) - data <- sc10x@data[,rnd] - labels <- labels[rnd] - } else { - data <- sc10x@data + + #if ((ncol(sc10x@data)>ds & ds!=0)){ + # rnd <- sample(1:ncol(sc10x@data),ds) + # data <- sc10x@data[,rnd] + # labels <- labels[rnd] + #} else { + # data <- sc10x@data + #} + #data <- as.data.frame(as.matrix(data)) + + cell.sample <- NULL + for (i in clusters){ + cell <- names(sc10x@ident[sc10x@ident==i]) + if (length(cell)>ds & ds!=0){ + rnd <- sample(1:length(cell),ds) + cell <- cell[rnd] + } + cell.sample <- c(cell.sample,cell) } + data <- sc10x@data[,colnames(sc10x@data) %in% cell.sample] data <- as.data.frame(as.matrix(data)) + labels <- labels[colnames(sc10x@data) %in% cell.sample] #Make labels for QuSAGE clust <- list() @@ -1071,37 +1113,13 @@ scCCA <- function(sc10x.1,sc10x.2,nm.1="D17",nm.2="D27",cc=FALSE){ } -sc3CCA <- function(sc10x.1,sc10x.2,sc10x.3,nm.1="D17",nm.2="D27",nm.3="D35",cc=FALSE,cca=25,acca=15){ - gc() - if (cc==TRUE){ - sc10x.1 <- ScaleData(object=sc10x.1,vars.to.regress=c("nUMI","percent.mito","S.Score","G2M.Score"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } else { - sc10x.1 <- ScaleData(object=sc10x.1,vars.to.regress=c("nUMI","percent.mito"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } - gc() - - gc() - if (cc==TRUE){ - sc10x.2 <- ScaleData(object=sc10x.2,vars.to.regress=c("nUMI","percent.mito","S.Score","G2M.Score"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } else { - sc10x.2 <- ScaleData(object=sc10x.2,vars.to.regress=c("nUMI","percent.mito"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } - gc() - - gc() - if (cc==TRUE){ - sc10x.3 <- ScaleData(object=sc10x.3,vars.to.regress=c("nUMI","percent.mito","S.Score","G2M.Score"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } else { - sc10x.3 <- ScaleData(object=sc10x.3,vars.to.regress=c("nUMI","percent.mito"),display.progress=FALSE,do.par=TRUE,num.cores=45) - } - gc() - - sc10x.1 <- FindVariableGenes(sc10x.1,do.plot=FALSE) - sc10x.2 <- FindVariableGenes(sc10x.2,do.plot=FALSE) - sc10x.3 <- FindVariableGenes(sc10x.3,do.plot=FALSE) - genes.hvg.1 <- head(rownames(sc10x.1@hvg.info),2000) - genes.hvg.2 <- head(rownames(sc10x.2@hvg.info),2000) - genes.hvg.3 <- head(rownames(sc10x.3@hvg.info),2000) +sc3CCA <- function(sc10x.1,sc10x.2,sc10x.3,nm.1="D17",nm.2="D27",nm.3="D35",cc=FALSE,cca=25,acca=15,lx=0.15,hx=3.5,ly=0.75){ + sc10x.1 <- FindVariableGenes(object=sc10x.1,mean.function=ExpMean,dispersion.function=LogVMR,x.low.cutoff=lx,x.high.cutoff=hx,y.cutoff=ly,do.plot=FALSE) + sc10x.2 <- FindVariableGenes(object=sc10x.2,mean.function=ExpMean,dispersion.function=LogVMR,x.low.cutoff=lx,x.high.cutoff=hx,y.cutoff=ly,do.plot=FALSE) + sc10x.3 <- FindVariableGenes(object=sc10x.3,mean.function=ExpMean,dispersion.function=LogVMR,x.low.cutoff=lx,x.high.cutoff=hx,y.cutoff=ly,do.plot=FALSE) + genes.hvg.1 <- sc10x.1@var.genes + genes.hvg.2 <- sc10x.2@var.genes + genes.hvg.3 <- sc10x.3@var.genes genes.hvg.Comb <- unique(c(genes.hvg.1,genes.hvg.2,genes.hvg.3)) genes.hvg.Comb <- intersect(genes.hvg.Comb,rownames(sc10x.1@scale.data)) genes.hvg.Comb <- intersect(genes.hvg.Comb,rownames(sc10x.2@scale.data)) @@ -1238,10 +1256,10 @@ scPseudotime <- function(sc10x,i,ds){ scMonocle <- reduceDimension(scMonocle,method='DDRTree') scMonocle <- orderCells(scMonocle) postscript(paste0("./analysis/pseudotime/Trajectory.State.eps"),paper="special",width=10,height=10,horizontal=FALSE) - plot_cell_trajectory(scMonocle,color_by="State") + plot(plot_cell_trajectory(scMonocle,color_by="State")) dev.off() postscript(paste0("./analysis/pseudotime/Trajectory.",i,".eps"),paper="special",width=10,height=10,horizontal=FALSE) - plot_cell_trajectory(scMonocle,color_by=i) + plot(plot_cell_trajectory(scMonocle,color_by=i)) dev.off() my_pseudotime_de <- differentialGeneTest(scMonocle,fullModelFormulaStr = "~sm.ns(Pseudotime)",cores=50) @@ -1249,17 +1267,17 @@ scPseudotime <- function(sc10x,i,ds){ my_pseudotime_gene <- my_pseudotime_gene$gene_short_name my_pseudotime_gene <- as.character(my_pseudotime_gene) postscript(paste0("./analysis/pseudotime/Genes.State.eps"),paper="special",width=10,height=10,horizontal=FALSE) - plot_genes_branched_pseudotime(scMonocle[my_pseudotime_gene,],color_by="State") + plot(plot_genes_branched_pseudotime(scMonocle[my_pseudotime_gene,],color_by="State")) dev.off() postscript(paste0("./analysis/pseudotime/Genes.",i,".eps"),paper="special",width=10,height=10,horizontal=FALSE) - plot_genes_branched_pseudotime(scMonocle[my_pseudotime_gene,],color_by=i) + plot(plot_genes_branched_pseudotime(scMonocle[my_pseudotime_gene,],color_by=i)) dev.off() my_pseudotime_de %>% arrange(qval) %>% head(50) %>% select(gene_short_name) -> gene_to_cluster gene_to_cluster <- gene_to_cluster$gene_short_name postscript(paste0("./analysis/pseudotime/Heatmap.eps"),paper="special",width=10,height=10,horizontal=FALSE) - my_pseudotime_cluster <- plot_pseudotime_heatmap(scMonocle[gene_to_cluster,],show_rownames=TRUE,return_heatmap=TRUE) + my_pseudotime_cluster <- plot_pseudotime_heatmap(scMonocle[gene_to_cluster,],show_rownames=TRUE,return_heatmap=TRUE) dev.off() #BEAM_res <- BEAM(scMonocle,branch_point=1,cores=50) diff --git a/r.scripts/sc-TissueMapper_RUN.Pd.R b/r.scripts/sc-TissueMapper_RUN.Pd.R index 295db929e35772d887c229f1db37813f555df44b..285435076c104a486dafec8c09290f7831bfef99 100644 --- a/r.scripts/sc-TissueMapper_RUN.Pd.R +++ b/r.scripts/sc-TissueMapper_RUN.Pd.R @@ -3,7 +3,7 @@ library(methods) library(optparse) library(Seurat) library(readr) -library(readxl) +#library(readxl) library(fBasics) library(pastecs) library(qusage) @@ -94,28 +94,28 @@ option_list=list( make_option("--p",action="store",default="DPrF",type='character',help="Project Name"), make_option("--g",action="store",default="ALL",type='character',help="Group To analyze"), make_option("--lg",action="store",default=500,type='integer',help="Threshold for cells with minimum genes"), - make_option("--hg",action="store",default=2500,type='integer',help="Threshold for cells with maximum genes"), + make_option("--hg",action="store",default=3000,type='integer',help="Threshold for cells with maximum genes"), make_option("--lm",action="store",default=0,type='numeric',help="Threshold for cells with minimum %mito genes"), make_option("--hm",action="store",default=0.1,type='numeric',help="Threshold for cells with maximum %mito genes"), - make_option("--lx",action="store",default=0.0125,type='numeric',help="x low threshold for hvg selection"), - make_option("--hx",action="store",default=3.5,type='numeric',help="x high threshold for hvg selection"), - make_option("--ly",action="store",default=0.5,type='numeric',help="y low threshold for hvg selection"), + make_option("--lx",action="store",default=0.2,type='numeric',help="x low threshold for hvg selection"), + make_option("--hx",action="store",default=5,type='numeric',help="x high threshold for hvg selection"), + make_option("--ly",action="store",default=1,type='numeric',help="y low threshold for hvg selection"), make_option("--cc",action="store",default=TRUE,type='logical',help="Scale cell cycle?"), make_option("--cca",action="store",default=50,type='integer',help="Number of CCAs to cacluate"), make_option("--acca",action="store",default=30,type='integer',help="Number of CCAs to align"), - make_option("--pc",action="store",default=25,type='integer',help="Number of PCs to cacluate"), - make_option("--hpc",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, pre-stress"), + make_option("--pc",action="store",default=50,type='integer',help="Number of PCs to cacluate"), + #make_option("--hpc",action="store",default=0.9,type='numeric',help="Max variance cutoff for PCs to use, pre-stress"), make_option("--res.prestress",action="store",default=1,type='numeric',help="Resolution to cluster, pre-stress"), make_option("--st",action="store",default=TRUE,type='logical',help="Remove stressed cells?"), - make_option("--stg",action="store",default="dws",type='character',help="Geneset to use for stress ID"), + make_option("--stg",action="store",default="go",type='character',help="Geneset to use for stress ID"), make_option("--cut.stress",action="store",default=0.9,type='numeric',help="Cutoff for stress score"), #make_option("--hpc.poststress",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, post-stress"), - make_option("--res.poststress",action="store",default=0.15,type='numeric',help="Resolution to cluster, post-stress"), - make_option("--ds",action="store",default=10000,type='integer',help="Number of cells to downsample"), + make_option("--res.poststress",action="store",default=0.2,type='numeric',help="Resolution to cluster, post-stress"), + #make_option("--ds",action="store",default=250,type='integer',help="Number of cells to downsample"), #make_option("--hpc.epi",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, Epi"), - make_option("--res.epi",action="store",default=0.15,type='numeric',help="Resolution to cluster, Epi"), + #make_option("--res.epi",action="store",default=0.2,type='numeric',help="Resolution to cluster, Epi"), #make_option("--hpc.st",action="store",default=0.85,type='numeric',help="Max variance cutoff for PCs to use, St"), - make_option("--res.st",action="store",default=0.15,type='numeric',help="Resolution to cluster, St"), + #make_option("--res.st",action="store",default=0.15,type='numeric',help="Resolution to cluster, St"), make_option("--cut.ne",action="store",default=0.999,type='numeric',help="Cutoff for NE score") ) opt=parse_args(OptionParser(option_list=option_list)) @@ -143,50 +143,68 @@ counts.cell.filtered <- results[[4]] counts.gene.filtered <- results[[5]] rm(results) +gc() +if (opt$cc==TRUE){ + sc10x <- ScaleData(object=sc10x,vars.to.regress=c("nUMI","percent.mito","S.Score","G2M.Score"),display.progress=FALSE,do.par=TRUE,num.cores=45) +} else { + sc10x <- ScaleData(object=sc10x,vars.to.regress=c("nUMI","percent.mito"),display.progress=FALSE,do.par=TRUE,num.cores=45) +} +gc() + sc10x.D17 <- scSubset(sc10x,"D17","D17") sc10x.D27 <- scSubset(sc10x,"D27","D27") sc10x.D35 <- scSubset(sc10x,"D35","D35") -results <- sc3CCA(sc10x.D17,sc10x.D27,sc10x.D35,"D17","D27","D35",cc=opt$cc,cca=opt$cca,acca=opt$acca) +results <- sc3CCA(sc10x.D17,sc10x.D27,sc10x.D35,"D17","D27","D35",cc=opt$cc,cca=opt$cca,acca=opt$acca,lx=opt$lx,hx=opt$hx,ly=opt$ly) sc10x <- results[[1]] genes.hvg.cca <- results[[2]] rm(results) -#results <- scPC(sc10xlx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="pre.stress",cca=TRUE) +rm(sc10x.D17) +rm(sc10x.D27) +rm(sc10x.D35) + +#results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="pre.stress",cca=TRUE) #sc10x <- results[[1]] #genes.hvg.prestress <- results[[2]] #pc.use.prestress <- results[[3]] #rm(results) sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=opt$res.prestress,folder="pre.stress",red="cca.aligned") +#sc10x <- scCluster(sc10x,pc.use=pc.use.prestress,res.use=opt$res.prestress,folder="pre.stress",red="pca") if (opt$st==TRUE){ - results <- scStress(sc10x,stg=opt$stg,res.use=opt$res.prestress,pc.use=pc.use.prestress,cut=opt$cut.stress) + results <- scStress(sc10x,stg=opt$stg,res.use=opt$res.prestress,cut=opt$cut.stress) sc10x <- results[[1]] counts.cell.filtered.stress <- results[[2]] sc10x.Stress <- results[[3]] rm(results) - #results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="post.stress",cca=TRUE) + #results <- scPC(sc10x,lx=opt$lx,hx=opt$hx,ly=opt$ly,cc=opt$cc,pc=opt$pc,hpc=opt$hpc,file="post.stress",cca=FALSE) #sc10x <- results[[1]] #genes.hvg.poststress <- results[[2]] #pc.use.poststress <- results[[3]] #rm(results) + #sc10x <- scCluster(sc10x,pc.use=pc.use.poststress,res.use=opt$res.poststress,folder="post.stress",red="pca") sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=opt$res.poststress,folder="post.stress",red="cca.aligned") + #sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=0.15,folder="post.stress",red="cca.aligned") + #sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=0.25,folder="post.stress",red="cca.aligned") + #sc10x <- scCluster(sc10x,pc.use=opt$acca,res.use=0.5,folder="post.stress",red="cca.aligned") } -gene.set1 <- read_delim("./genesets/DEG_FMSt_5FC.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=FALSE) -gene.set1 <- as.list(gene.set1) -names(gene.set1) <- "St" -gene.set <- c(gene.set1) gene.set1 <- read_delim("./genesets/DEG_Epi_5FC.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=FALSE) gene.set1 <- as.list(gene.set1) names(gene.set1) <- "Epi" +gene.set <- c(gene.set1) +gene.set1 <- read_delim("./genesets/DEG_FMSt_5FC.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=FALSE) +gene.set1 <- as.list(gene.set1) +names(gene.set1) <- "St" gene.set <- c(gene.set,gene.set1) rm(gene.set1) gc() -results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=opt$ds,nm="Lin",folder="lin") +min.all <- min(table(sc10x@meta.data[,paste0("res",opt$res.poststress)])) +results <- scQuSAGE(sc10x,gs=gene.set,res.use=opt$res.poststress,ds=min.all,nm="Lin",folder="lin") sc10x <- results[[1]] results.cor.Lin <- results[[2]] results.clust.Lin.id <- results[[3]] @@ -199,8 +217,27 @@ if (any(levels(sc10x@ident)=="Unknown")){ } else { sc10x.St <- scSubset(sc10x,i="Lin",g="St") } +sc10x.Epi <- SetAllIdent(object=sc10x.Epi,id=paste0("res",opt$res.poststress)) +sc10x.Epi <- BuildClusterTree(sc10x.Epi,do.reorder=TRUE,reorder.numeric=TRUE,do.plot=FALSE) +sc10x.Epi <- StashIdent(object=sc10x.Epi,save.name=paste0("res",opt$res.poststress)) +sc10x.St <- SetAllIdent(object=sc10x.St,id=paste0("res",opt$res.poststress)) +sc10x.St <- BuildClusterTree(sc10x.St,do.reorder=TRUE,reorder.numeric=TRUE,do.plot=FALSE) +sc10x.St <- StashIdent(object=sc10x.St,save.name=paste0("res",opt$res.poststress)) -sc10x.Epi <- scCluster(sc10x.Epi,pc.use=opt$acca,res.use=opt$res.epi,folder="epi",red="cca.aligned") +sc10x.Epi <- RunTSNE(object=sc10x.Epi,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) +postscript(paste0("./analysis/tSNE/epi/tSNE_Sample.eps")) +plot <- TSNEPlot(object=sc10x.Epi,group.by="samples",pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) +plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) +plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) +plot(plot) +dev.off() +postscript(paste0("./analysis/tSNE/epi/tSNE_res",opt$res.poststress,".eps")) +plot <- TSNEPlot(object=sc10x.Epi,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) +plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) +plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) +plot(plot) +dev.off() +rm(plot) gene.set1 <- read_delim("./genesets/DEG_BE_5FC.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=FALSE) gene.set1 <- as.list(gene.set1) @@ -216,14 +253,15 @@ names(gene.set1) <- "OE" gene.set <- c(gene.set,gene.set1) rm(gene.set1) gc() -results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=0.15,ds=opt$ds,nm="Epi.dws",folder="epi") +min.epi <- min(table(sc10x.Epi@meta.data[,paste0("res",opt$res.poststress)])) +results <- scQuSAGE(sc10x.Epi,gs=gene.set,res.use=opt$res.poststress,ds=min.epi,nm="Epi.dws",folder="epi") sc10x.Epi <- results[[1]] results.cor.Epi.dws <- results[[2]] results.clust.Epi.dws.id <- results[[3]] rm(results) rm(gene.set) -sc10x.Epi <- SetAllIdent(object=sc10x.Epi,id=paste0("res",opt$res.epi)) +sc10x.Epi <- SetAllIdent(object=sc10x.Epi,id=paste0("res",opt$res.poststress)) OE <- levels(factor(sc10x.Epi@ident[sc10x.Epi@meta.data$Epi.dws=="OE"])) OE.cells <- NULL for (i in 1:length(OE)){ @@ -243,6 +281,7 @@ dev.off() rm(plot) rm(OE) rm(OE.cells) +rm(i) gene.set1 <- read_csv("./genesets/Basal cells-signature-genes.csv") gene.set1 <- gene.set1[,2] @@ -261,11 +300,71 @@ names(gene.set1) <- "CGC" gene.set<- c(gene.set,gene.set1) rm(gene.set1) gc() -results.cor.Epi.lgea <- scQuSAGEsm(sc10x.Epi,gs=gene.set,ds=opt$ds,nm="Epi.dws.sub",folder="lgea") +results.cor.Epi.lgea <- scQuSAGEsm(sc10x.Epi,gs=gene.set,ds=min.epi,nm="Epi.dws.sub",folder="lgea") rm(gene.set) +#sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=opt$res.st,folder="st",red="cca.aligned") +#sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=0.2,folder="st",red="cca.aligned") +#sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=0.25,folder="st",red="cca.aligned") +#sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=0.5,folder="st",red="cca.aligned") +#sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=0.1,folder="st",red="cca.aligned") + +sc10x.St <- RunTSNE(object=sc10x.St,reduction.use="cca.aligned",dims.use=1:opt$acca,do.fast=TRUE) +postscript(paste0("./analysis/tSNE/st/tSNE_Sample.eps")) +plot <- TSNEPlot(object=sc10x.St,group.by="samples",pt.size=2.5,do.return=TRUE,vector.friendly=FALSE) +plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) +plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) +plot(plot) +dev.off() +postscript(paste0("./analysis/tSNE/st/tSNE_res",opt$res.poststress,".eps")) +plot <- TSNEPlot(object=sc10x.St,pt.size=5,do.label=TRUE,label.size=10,do.return=TRUE,vector.friendly=FALSE) +plot <- plot+theme(axis.text.x=element_text(size=20),axis.text.y=element_text(size=20),axis.title.x=element_text(size=20),axis.title.y=element_text(size=20),legend.text=element_text(size=20)) +plot <- plot+guides(colour=guide_legend(override.aes=list(size=10))) +plot(plot) +dev.off() +rm(plot) + +gene.set1 <- read_delim("./genesets/DEG_C5.BP.M11704.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) +gene.set1 <- as.list(gene.set1[-1,]) +names(gene.set1) <- "Endo" +gene.set <- c(gene.set1) +gene.set1 <- read_delim("./genesets/DEG_C5.BP.M10794.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) +gene.set1 <- as.list(gene.set1[-1,]) +names(gene.set1) <- "SM" +gene.set <- c(gene.set,gene.set1) +gene.set1 <- read_delim("./genesets/DEG_C5.BP.M13024.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) +gene.set1 <- as.list(gene.set1[-1,]) +names(gene.set1) <- "Fib" +gene.set <- c(gene.set,gene.set1) +gene.set1 <- read_delim("./genesets/DEG_C5.BP.M10124.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) +gene.set1 <- as.list(gene.set1[-1,]) +names(gene.set1) <- "Leu" +gene.set <- c(gene.set,gene.set1) +rm(gene.set1) +gc() +min.st <- min(table(sc10x.Epi@meta.data[,paste0("res",opt$res.poststress)])) +results <- scQuSAGE(sc10x.St,gs=gene.set,res.use=0.2,ds=min.st,nm="St.go",folder="st") +sc10x.St <- results[[1]] +results.cor.St.go <- results[[2]] +results.clust.St.go.id <- results[[3]] +rm(results) +rm(gene.set) + +sc10x.Epi.NE <- scNE(sc10x.Epi,neg="EurUro",cut=opt$cut.ne) + +sc10x.Epi <- scMergeSubClust(sc10x.Epi,i="Epi.dws.sub",g=c("BE","LE"),nm="Merge") + +sc10x.St <- scMergeSubClust(sc10x.St,i="St.go",g=c("Endo","SM","Fib","Leu"),nm="Merge") + +sc10x <- scMerge(sc10x,sc10x.Epi,sc10x.St,i.1="Merge",i.2="Merge",nm="Merge_Epi.dws_St.go") + +sc10x <- scMerge(sc10x,sc10x,sc10x.Epi.NE,i.1="Merge_Epi.dws_St.go",i.2="NE",nm="Merge_Epi.dws_St.go_NE") + +sc10x.Epi <- scMerge(sc10x.Epi,sc10x.Epi,sc10x.Epi.NE,i.1="Epi.dws.sub",i.2="NE",nm="Epi.dws.sub_NE") + gene.orthog <- read.delim("./genesets/Ensemble.mus-hum.txt") -gene.set1 <- read_excel("./genesets/SupTab3_Consensus_Sigs.xlsx",skip=5) +#gene.set1 <- read_excel("./genesets/SupTab3_Consensus_Sigs.xlsx",skip=5) +gene.set1 <- read_csv("./genesets/SupTab3_Consensus_Sigs.csv",skip=6) gene.set2 <- as.data.frame(gene.set1$Basal[!is.na(gene.set1$Basal)]) colnames(gene.set2) <- "genes" gene.set2 <- as.data.frame(merge(gene.set2,gene.orthog,by.x="genes",by.y="Gene.name")[,4]) @@ -303,7 +402,8 @@ gene.set2 <- as.list(gene.set2) names(gene.set2) <- "Ionocyte" gene.set <- c(gene.set,gene.set2) rm(gene.set2) -gene.set1 <- read_excel("./genesets/SupTab6_Krt13_Hillock.xlsx",skip=5) +#gene.set1 <- read_excel("./genesets/SupTab6_Krt13_Hillock.xlsx",skip=5) +gene.set1 <- read_csv("./genesets/SupTab6_Krt13_Hillock.csv",skip=6) gene.set1 <- gene.set1[gene.set1$FDR<=0.05 & gene.set1$'log2 fold-change (MAST)'>=1.5,1] colnames(gene.set1) <- "genes" gene.set1 <- as.data.frame(merge(gene.set1,gene.orthog,by.x="genes",by.y="Gene.name")[,4]) @@ -313,48 +413,11 @@ gene.set <- c(gene.set,gene.set1) rm(gene.set1) rm(gene.orthog) gene.set <- lapply(gene.set,droplevels) -results.cor.Epi.MusLungHierarchy <- scQuSAGEsm(sc10x.Epi,gs=gene.set,ds=opt$ds,nm="Epi.dws.sub",folder="MusLungHierarchy") -rm(gene.set) - -sc10x.St <- scCluster(sc10x.St,pc.use=opt$acca,res.use=opt$res.st,folder="st",red="cca.aligned") - -gene.set1 <- read_delim("./genesets/DEG_C5.BP.M11704.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- as.list(gene.set1[-1,]) -names(gene.set1) <- "Endo" -gene.set <- c(gene.set1) -gene.set1 <- read_delim("./genesets/DEG_C5.BP.M10794.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- as.list(gene.set1[-1,]) -names(gene.set1) <- "SM" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/DEG_C5.BP.M13024.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- as.list(gene.set1[-1,]) -names(gene.set1) <- "Fib" -gene.set <- c(gene.set,gene.set1) -gene.set1 <- read_delim("./genesets/DEG_C5.BP.M10124.txt","\t",escape_double=FALSE,trim_ws=TRUE,col_names=TRUE) -gene.set1 <- as.list(gene.set1[-1,]) -names(gene.set1) <- "Leu" -gene.set <- c(gene.set,gene.set1) -rm(gene.set1) -gc() -results <- scQuSAGE(sc10x.St,gs=gene.set,res.use=0.15,ds=opt$ds,nm="St.go",folder="st") -sc10x.St <- results[[1]] -results.cor.St.go <- results[[2]] -results.clust.St.go.id <- results[[3]] -rm(results) +results.cor.Epi.MusLungHierarchy <- scQuSAGEsm(sc10x.Epi,gs=gene.set,ds=min.epi,nm="Epi.dws.sub_NE",folder="MusLungHierarchy") rm(gene.set) -sc10x.Epi.NE <- scNE(sc10x.Epi,neg="EurUro",cut=opt$cut.ne) - -sc10x.Epi <- scMergeSubClust(sc10x.Epi,i="Epi.dws.sub",g=c("BE","LE"),nm="Merge") - -sc10x.St <- scMergeSubClust(sc10x.St,i="St.go",g=c("Endo","SM","Fib","Leu"),nm="Merge") - -sc10x <- scMerge(sc10x,sc10x.Epi,sc10x.St,i.1="Merge",i.2="Merge",nm="Merge_Epi.dws_St.go") - -sc10x <- scMerge(sc10x,sc10x,sc10x.Epi.NE,i.1="Merge_Epi.dws_St.go",i.2="NE",nm="Merge_Epi.dws_St.go_NE") - gene.set.c2.all <- read.gmt("./genesets/c2.all.v6.1.symbols.gmt") -results.cor.c2.ALL <- scQuSAGElg(sc10x,gs=gene.set.c2.all,ds=opt$ds,nm="Merge_Epi.dws_St.go",folder="c2.all") +results.cor.c2.ALL <- scQuSAGElg(sc10x,gs=gene.set.c2.all,ds=min.all,nm="Merge_Epi.dws_St.go",folder="c2.all") rm(gene.set.c2.all) sc10x <- SetAllIdent(object=sc10x,id="Merge_Epi.dws_St.go") @@ -476,6 +539,7 @@ for (g in c("D17","D27","D35")){ } sctSNEHighlight(sc10x,i="Pz",g="Pz",file="D") sctSNEHighlight(sc10x,i="Tz",g="Tz",file="D") +rm(i) rm(g) scCustHeatmap(sc10x.Epi,i="Epi.dws.sub",gs=c(genes.deg.BE.unique,genes.deg.LE.unique,genes.deg.OE1.unique,genes.deg.OE2.unique),g=c("BE","LE","OE1","OE2"))