GSM6630613: H1-0_siRNA_Input; Mus musculus; ChIP-Seq
H1.0 and FLAG ChIP-seq: Primary isolated murine cardiac fibroblasts were transfected with Adv-GFP-H1.0-FLAG for 48 hours.  FLAG and H1.0 chromatin immunoprecipitation was performed using anti-FLAG (Sigma, cat. no. F1804) and anti-H1.0 (Proteintech, cat. no. 17510-1-AP) antibodies. Chromatin shearing was performed using the truChIP Chromatin Shearing kit (Covaris, cat. no. 520154) according to the manufacturer's instructions. DNA fragment size was assessed using a TapeStation 4200 (Agilent). Samples in the 300-500bp range were used for immunoprecipitation using the ChIP-IT High Sensitivity kit (Active Motif, cat. no. 53040). DNA was purified using a Zymo DNA Clean & Concentrator-5 kit (Zymo, cat. no. D4014). Library preparation and DNA sequencing were performed by the UCLA Technology Center for Genomics & Bioinformatics Core. Approximately 35 million paired-end reads per sample (2x150bp) were generated and used for bioinformatic analyses. H3K27ac ChIP-seq: Three biological replicates of isolated cardiac fibroblasts from passage 1 were transfected with H1.0 or scrambled siRNAs for 48 hours and treated with TGF-β (10 ng/mL) for 24 hours. H3K27ac immunoprecipitation was performed using the same experimental strategy as the H1.0 ChIP-seq experiment, but using an anti-H3K27ac antibody (Abcam, cat. no. ab4729). For each biological condition, a combined input sample of sonicated genomic DNA was obtained from all three biological replicates. Library preparation and paired-end sequencing were performed at the UCLA Technology Center for Genomics and Bioinformatics Core, resulting in ~50-70 million read pairs (2x150bp) per sample.