The DNA binding of R270C mutant form of p53 in mouse osteosarcoma AXT cells

Mouse osteosarcoma AXT cells were established from bone marrow stromal cells of Ink4a / Arf-null mice by overexpressing human c-MYC (Oncogene 29: 5687-5699, 2010, Cancer Research 74: 6531-6541, 2014). AXT cells harbor R270C mutant form of p53 (equivalent to human R273C mutant). R270C mutant p53 did not show transcriptional activity nor exert critical roles in osteosarcoma progression in vivo. Therfore, we examined whether mutant p53 could bind to chromatin by ChIP-seq analysis using AXT cells. The ChIP-seq of H3K4me3 and H3K9me3 was also performed as the marker of transcriptional activation and repression, respectively. Overall design: Chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for mouse p53 as well as the histone modifications H3K4me3 and H3K9me3 in AXT cells cultured in 10% FBS containing IMDM medium.