Stress-induced signaling to the nucleosome in serum deprived mouse fibroblasts.. Mus musculus

We have employed chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq) to analyze changes in chromatin architecture as well as the occupancy of two RNA Polymerase II (RNAPII) isoforms, initiation-competent (RNAPIIS5ph) as well as elongation-competent (RNAPIIS2ph) upon stress induction. We used resting mouse Swiss 3T3 fibroblasts, either untreated (control) or treated with anisomycin to induce the p38/MAP kinase pathway. Overall design: Serum starved (72 h 0.2% FCS) mouse 3T3 cells were treated with anisomycin (188.5 nM) for 1 h (in duplicates). Untreated, serum-starved cells were used as a control. Isolated chromatin was subjected to immunoprecipitation with the following antibodies: a-H3S28ph, a-H3K9ac, a-H3K4me3, a-RNAPIIS5ph and a-RNAPIIS2ph. Resulting DNA was sequenced using Illumina platforms.