Fig. 7

Transcriptome analysis upon transient CD271 overexpression. a RNAseq of cells before, during and after CD271 transient overexpression was performed and analyzed using edgeR. Genes were filtered for Log2 ≥ +0.27 and ≤−0.27, p-value < 0.05, FDR < 0.05, which resulted in 160 genes significantly upregulated upon CD271 overexpression (comparing CMVTOCD271 + DOX 24 h over CMVTOCD271 –DOX24 h) with subsequent significant downregulation (comparing CMVTOCD271 + DOX 24 h/−DOX 48 h over CMVTOCD271 + DOX 24 h) and vice versa in 303 genes significantly downregulated upon CD271 overexpression with subsequent significant upregulation. Heatmap represents row z-scores of normalized counts from RNAseq. b Top 20 gene ontology process enrichments from MetaCore^TM. Numbers in brackets indicate the number of differentially expressed genes above the total number of genes within an annotated pathway. c Selected “switching” genes from a involved in cholesterol and lipid biosynthesis pathways. Again, heatmap represents row z-scores of normalized counts from RNA Seq. d Survival curves based on TCGA data for expression of cholesterol biosynthesis genes in human melanoma samples. Data from 114 patients per condition were analyzed, which represent 25% of highest and lowest expressing patients. e Selected “switching” genes from a involved in cell cycle regulation. Again, heatmap represents row z-scores of normalized counts from RNA Seq. f qRT-PCR for cell cycle genes in ICD overexpressing and EV melanoma cells. g qRT-PCR for cholesterol and lipid biosynthesis genes in ICD overexpressing and EV melanoma cells. For f and g, n = 3 for each condition. P values > 0.05 = n.s., P values ≤ 0.05 = *, P values ≤ 0.01 = **, P values ≤ 0.001 = ***. Error bars for f and g indicate S.D. All experiments done with cell line M010817