ChIP Assays for Histone Modifications

ChIP assays were performed as previously described [73 (link)]. The antibodies used for immunoprecipitation were: normal rabbit IgG (Santa Cruz), polyclonal antibody for PARP-1 (Enzo Lifescience), H3Ac (Millipore), H3K4me3 (Millipore) and H3K27me3 (Millipore). The sense oligonucleotide used to amplify the promoter region was 5′-AACCCCGCCTCGGAGGAGT-3′ and the antisense oligonucleotide was 5′-CCAATCGGAGGCTCGTCT-3′. Real-Time PCR assays were carried out to amplify the enhancer region as already described [62 (link)]. The sense oligonucleotide used was FW 5′-GAGCAGCCCTTAATGACTTG-3′ and the antisense oligonucleotide was 5′-CCCAACTCCCTAACTTCCC-3′. The sense oligonucleotide used to amplify the promoter region of ITPR1 was 5′-ACTGAGGTCGCGGTTTGTAT-3′ and the antisense oligonucleotide was 5′-AAGGAGCCGTGTTGTGACTT-3′ [63 (link)].