## ----maits_load, message=FALSE, warning=FALSE------------------------------ suppressPackageStartupMessages({ library(MAST) library(singleCellTK) library(xtable) }) data(maits, package="MAST") maits_sce <- createSCE(assayFile = t(maits$expressionmat), annotFile = maits$cdat, featureFile = maits$fdat, assayName = "logtpm", inputDataFrames = TRUE, createLogCounts = FALSE) rm(maits) ## ----maits_summarize, results='asis'--------------------------------------- knitr::kable(summarizeTable(maits_sce, useAssay = "logtpm")) ## ----maits_colnames-------------------------------------------------------- colnames(colData(maits_sce)) table(colData(maits_sce)$ourfilter) ## ----maits_filter---------------------------------------------------------- maits_subset <- maits_sce[, colData(maits_sce)$ourfilter] table(colData(maits_subset)$ourfilter) ## ----maits_filter_table, results='asis'------------------------------------ knitr::kable(summarizeTable(maits_subset, useAssay = "logtpm")) ## ----maits_availablereduceddims-------------------------------------------- reducedDims(maits_subset) ## ----maits_getpcatsne------------------------------------------------------ maits_subset <- getPCA(maits_subset, useAssay = "logtpm", reducedDimName = "PCA_logtpm") maits_subset <- getTSNE(maits_subset, useAssay = "logtpm", reducedDimName = "TSNE_logtpm") reducedDims(maits_subset) ## ----maits_pca------------------------------------------------------------- plotPCA(maits_subset, reducedDimName = "PCA_logtpm", colorBy = "condition") ## ----maits_tsne------------------------------------------------------------ plotTSNE(maits_subset, reducedDimName = "TSNE_logtpm", colorBy = "condition") ## ----maits_convert_symbols, message=FALSE---------------------------------- suppressPackageStartupMessages({ library(org.Hs.eg.db) }) maits_entrez <- maits_subset maits_subset <- convertGeneIDs(maits_subset, inSymbol = "ENTREZID", outSymbol = "SYMBOL", database = "org.Hs.eg.db") #to remove confusion for MAST about the gene name: rowData(maits_subset)$primerid <- NULL ## ----maits_thresh, fig.height=8, message=FALSE----------------------------- thresholds <- thresholdGenes(maits_subset, useAssay = "logtpm") par(mfrow = c(5, 4)) plot(thresholds) par(mfrow = c(1, 1)) ## ----maits_MAST, message=FALSE--------------------------------------------- mast_results <- MAST(maits_subset, condition = "condition", useThresh = TRUE, useAssay = "logtpm") ## ----maits_violin, fig.height=8, message=FALSE----------------------------- MASTviolin(maits_subset, useAssay = "logtpm", fcHurdleSig = mast_results, threshP = TRUE, condition = "condition") ## ----maits_lm, fig.height=8, message=FALSE--------------------------------- MASTregression(maits_subset, useAssay = "logtpm", fcHurdleSig = mast_results, threshP = TRUE, condition = "condition") ## ----maits_heatmap--------------------------------------------------------- plotDiffEx(maits_subset, useAssay = "logtpm", condition = "condition", geneList = mast_results$Gene[1:100], annotationColors = "auto", displayRowLabels = FALSE, displayColumnLabels = FALSE) ## ----maits_gsva, message=FALSE--------------------------------------------- gsvaRes <- gsvaSCE(maits_entrez, useAssay = "logtpm", "MSigDB c2 (Human, Entrez ID only)", c("KEGG_PROTEASOME", "REACTOME_VIF_MEDIATED_DEGRADATION_OF_APOBEC3G", "REACTOME_P53_INDEPENDENT_DNA_DAMAGE_RESPONSE", "BIOCARTA_PROTEASOME_PATHWAY", "REACTOME_METABOLISM_OF_AMINO_ACIDS", "REACTOME_REGULATION_OF_ORNITHINE_DECARBOXYLASE", "REACTOME_CYTOSOLIC_TRNA_AMINOACYLATION", "REACTOME_STABILIZATION_OF_P53", "REACTOME_SCF_BETA_TRCP_MEDIATED_DEGRADATION_OF_EMI1"), parallel.sz=1) set.seed(1234) gsvaPlot(maits_subset, gsvaRes, "Violin", "condition") gsvaPlot(maits_subset, gsvaRes, "Heatmap", "condition") ## ----load_bladderbatch, message=FALSE-------------------------------------- library(bladderbatch) data(bladderdata) dat <- bladderEset pheno <- pData(dat) edata <- exprs(dat) bladder_sctke <- createSCE(assayFile = edata, annotFile = pheno, assayName = "microarray", inputDataFrames = TRUE, createLogCounts = FALSE) ## ----plot_var_microarray, message=FALSE------------------------------------ plotBatchVariance(bladder_sctke, useAssay="microarray", batch="batch", condition = "cancer") ## ----run_combat, message=FALSE--------------------------------------------- assay(bladder_sctke, "combat") <- ComBatSCE(inSCE = bladder_sctke, batch = "batch", useAssay = "microarray", covariates = "cancer") ## ----plot_var_postcombat, message=FALSE------------------------------------ plotBatchVariance(bladder_sctke, useAssay="combat", batch="batch", condition = "cancer") ## ----sessionInfo, echo=FALSE----------------------------------------------- sessionInfo()