Fig. 4: p63 is a pioneer factor essential for the establishment of an LCE2 enhancer. | Cell Death Discovery

Fig. 4: p63 is a pioneer factor essential for the establishment of an LCE2 enhancer.

From: Epigenetic priming of an epithelial enhancer by p63 and CTCF controls expression of a skin-restricted gene XP33

Fig. 4

A ChIP-seq signal tracks for p63 in human keratinocytes induced for 2, 4 or 7 days of differentiation. B ChIP-seq signal tracks for p63 in H3K27ac in human fibroblasts expressing either empty vector or ΔNp63α. At the bottom, ChIP-seq signal tracks for H3K27ac in human fibroblasts keratinocytes. C RNA-seq signal track of fibroblast expressing either empty vector, or wild-type ΔNp63α, or DND-binding ___domain mutant of ΔNp63α. D qPCR analysis of XP33 expression in Ker-CT was silenced for p63 and induced to differentiate for 9 days. Western blot confirms an efficient knock-down. n = 4 (biological replicates), P by one-way ANOVA. E Co-immunoprecipitation in HEK293 overexpressing ΔNp63α-HA using either anti-CTCF or anti-IgG antibodies. Western blot for HA and CTCF. n = 2 (biological replicates). F (Left) Schematic shows strategy to select differentiation-specific skin-restricted TFs which may regulate the expression of XP33. (Right) Dot plot showing the distribution of human TFs based on the specificity of expression in skin and correlation of their expression with levels of XP33. Top hits are shown in a beige box. G (Top) ChIP-seq signals for KLF4 and ZNF750 in differentiated keratinocytes. The enlarged area of the LCE2 locus is shown on the right. (Bottom) qPCR analysis of XP33 expression in Ker-CT silenced for KLF4 or ZNF750 and induced to differentiate for 9 days. Western blot confirms an efficient knock-down. n = 3 (biological replicates), P by one-way ANOVA.

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