Profiling Epidermal Differentiation via ChIP-seq

Human primary keratinocytes under proliferating, early, mid-, and late differentiation condition generated from HKC1 were used for ChIP and ChIP-seq analysis. ChIP-sequencing datasets were generated with an antibody recognizing the alpha isoforms of p63 (H129, Santa Cruz), an antibody that detects all forms of the large subunit of RNA polymerase II (8WG16, Santa Cruz), and a H3K27ac antibody recognizing the acetylation of the lysine 27 residue of histone H3 (H3K27ac, C15410174, pAb-174-050, Diagenode). ChIP-qPCR experiments were performed using 2 μg RUNX1 antibody (ab23980, Abcam) and 1 μg TFAP2 antibody (C18X, Santa cruz). RNAPII, p63, H3K27ac, RUNX1, and TFAP2 ChIP experiments were performed as previously described 65 (link), with a minor change of using magnetic beads (Novex by Life Technologies, 10008D and 10009D).