Chromatin Immunoprecipitation of HPV-31

Chromatin immunoprecipitations were performed using CIN612 or stable HPV-31 (WT or L2 Mutant 3X) cells grown as described above. Before fixation, cells were treated with versene to remove J2 fibroblast feeders. Cells were then seeded and grown to 80% confluency. Chromatin-immunoprecipitation was performed as previously described using SMC1 (Abcam, Cat. #ab9262), γ-H2AX (Millipore Cat. #05–636), CTCF (Millipore, Cat. #07–729), Normal Rabbit IgG (Santa Cruz, Cat. #SC-2027), Normal Mouse IgG Antibody (Santa Cruz, Cat. #SC-2025), Normal Mouse IgG (Millipore, Cat. #12-371B) and Protein-G Dynabeads (Invitrogen Cat. #100-03D) [50 (link)]. Real-time, touchdown PCR was performed with the Lightcycler 480 (Roche) and the HPV-31 CTCF Trio Primers and URR Primers; Forward: 5’-TTTGGTGGGTTGGGTATTGG-3’, Reverse:5’-GTAGGAGGCTGCAATACAGATG-3’. Forward, 5’-AACTGCCAAGGTTGTGTCATGC 3’, Reverse, 5’-TGGCGTCTGTAGGTTTGCAC-3’.