Fig. 4

UTX suppresses AP-1 components and gliogenesis. a ATAC-seq datasets from H7 and H9 UTX-WT-differentiating cells were compared to those of H7 and H9 UTX-KO-differentiating cells. b Enrichment of consensus motifs in nucleosome-free regions (from ATAC-seq datasets) that significantly increased signals in UTX-KO vs. UTX-WT differentiating cells. GSEA showed significant enrichment of AP-1 target genes in c upregulated genes and d in distal elements with increased ATAC-seq signals in UTX-KO (vs. WT) cell differentiation. e At the JUN and JUNB loci, UTX ChIP-seq profiles in hNSCs and pS2-RNApol2 CUT&RUN-seq in wild-type and UTX-KO-differentiating cells. f Relative levels of JUN and FOS genes (normalized by counts per million from RNA-seq datasets) in UTX-WT and UTX-KO cells at neuron time point of differentiation. g Volcano plot of pS2-RNApol2 CUT&RUN-seq signals in UTX-KO/WT differentiating cells. fc, fold change. h Analysis of the transcripts of glial/astrocytic markers S100B and GFAP expressed by UTX-KO cells that were treated by DMSO or AP-1 inhibitors during neural differentiation. Observations were made with 3 biological replicates. We used 15 μM of SR-11302 and 30 μM of T-5224 at concentrations similar to those used by previous studies [48,49,50]. i GFAP immunofluorescence of UTX-KO differentiating cells that were treated by DMSO or AP-1 inhibitors. Bar, 100 μm. Quantification of GFAP-positive cells in the 6 assayed groups. More than 250 cells were counted. ns indicates not significant. * and ** indicate P < 0.05 and 0.01 by two-side Student’s t test