ChIP-qPCR Assay for p63 Regulation

ChIP-qPCR assays were performed on murine embryonic fibroblasts isolated from p63 flox/flox (f/f) mice (gift of Dr. Elsa Flores; Moffitt Cancer Center & Research Institute), as previously described^{8 (link)}. Briefly, cells were seeded onto 150 mm dishes at density of 10⁶ cells/dish, and treated 24 h later with adenoviral vectors expressing GFP or Cre recombinase (AdGFP and AdCre; 150 moi). These cells were collected seven days later and processed using ChromaFlash Chromatin Extraction kits (p-2001; Epigenetek) per manufacturer’s protocol. ChIP was then conducted using H3K27Ac antibody (ab4729; Abcam), HDAC1 antibody (815104; Biolegend) or normal rabbit immunoglobulin G (AB-105-C; R&D systems), as previously described^{27 (link)}. ChIP-qPCR was performed using primers synthesized by Sigma Aldrich (Supplemental Table S2). Fold-enrichment of PCR products was calculated after normalization with input of all three types of infected cells.