Fig. 4

AF10 expression maintains somatic cell identity. a Sample distance matrix of RNA-sequencing replicate samples. b Gene set enrichment analysis (GSEA) of transcriptome data of sgAF10 cells with respect to pluripotency-related and fibroblast-related gene sets. NES: normalized enrichment score, q: false discovery rate (FDR) q-value. c Gene set enrichment analysis (GSEA) of transcriptome data of sgAF10 cells with respect to iDOT1L_DOWN and iDOT1L_UP gene sets. NES: normalized enrichment score, q: false discovery rate (FDR) q-value. d mRNA levels for a set of DOT1L-regulated genes in AF10 knock-out fibroblasts as determined by qRT-PCR. β-actin was used as an internal control. Gene expression levels were normalized to sgControl expressing fibroblasts for sgAF10 samples and DMSO treated cells for iDOT1L samples. Two biological replicates are indicated for sgAF10 samples and bar graph indicates the average of replicates. e Fold change in the number of Tra-1-60-positive colonies derived from AF10 sgRNA expressing cells in the presence of DMSO or a DOT1L inhibitor (iDOT1L; EPZ004777). P values were 0.001 for sgAF10-1 and 0.004 for sgAF10-2. n.s. not significant. f Fold change in the number of Tra-1-60-positive colonies derived from control or AF10 knock-down cells (shAF10) treated with either vehicle (DMSO) or iDOT1L (EPZ004777). P values were 0.034 for shAF10-1 and 0.007 for shAF10-2. n.s., not significant. g Immunoblot for H3K79me2 in double KO cells. Total H3 levels were used as loading control. Replicate of this experiment in Additional file 1: Figure S3c. Quantifications were normalized to gNT sample. h Fold change in the number of Tra-1-60-positive colonies derived from double knock-out cells expressing DOT1L and AF10 targeting sgRNAs. P values were 0.031 for sgAF10-1 and 0.014 for sgAF10-2. n.s. not significant