Fig. 5: The expression of TGFβ-stimulated poised genes is highly dependent on PHF13.

A Aggregate profiles show the occupancy of H3K4me3, H3K27me3, and H3K27ac surrounding the center of PHF13-occupied poised chromatin regions (±5 kb) in siNC, siNC + TGFβ (NCT), and siPHF13 + TGFβ (PHFT) Panc-1 cells. P values were calculated by Wilcoxon–Mann–Whitney Test. ns nonsignificant difference; **0.01 < p < 0.001; ***p < 0.001. B Heatmaps show gene expression of TGFβ-activated poised genes in siNC (NC), siPHF13 (PHF), siNC + TGFβ (NCT), and siPHF13 + TGFβ (PHFT) Panc1 cells. The color scale bar shows the row-wise normalized expression signal. C GO analysis of the TGFβ-activated poised genes. The top 10 significantly enriched gene terms were shown. Western blot analysis of nuclear extract inputs, anti-PHF13 IP (D), anti-EZH2 IP (E), and anti-IgG IP from Panc1 nuclear extract. F Western blot showing the level of H3K27me3 in the given samples. The protein TBP was set as a loading control. The down numbers indicated the normalized signal of H3K27me3. The signal of each band was measured using ImageJ. The signal of H3K27me3 was normalized to TBP and represented as fold change relative to the control cells. G Genome browser tracks the normalized signal of chromatin assembly (ATAC-seq), EZH2, PHF13, H3K27me3, H3K4me3 occupancy, and mRNA-seq data at PCDHB8 and PAPLN locus from siNC (NC), siPHF13 (PHF), siNC + TGFβ (NCT), siPHF13 + TGFβ (PHFT) Panc1 cells. H ChIP-qPCR analysis of H3K27me3 occupancy near the TSS of the PCDHB8 and PAPLN genes in siControl treated, PHF13-depleted, TGFβ-treated, and PHF13-depleted TGFβ-treated Panc-1 cells. p value was calculated using a two-side t test. *p < 0.05; ***p < 0.001. ChIP-qPCR for IgG was set as a negative control. The dotted line indicated the average signal of IgG.