GSM5834957: 6_Uninjured_H3K27me3; Rattus norvegicus; ChIP-Seq
Tissue collection was performed at 7 days after balloon angioplasty. From the same animal, we collected the endothelial-damaged central segment of the balloon-injured common carotid artery (denoted as “injured” throughout this study) and the contralateral common carotid artery without angioplasty (sham control, denoted as “uninjured”). To preserve the artery “real-time” epigenetic information, the collected tissue samples were immediately snap-frozen in liquid N2. Artery tissues (injured or uninjured) from 50 rats were pooled for ChIPseq analysis at Active Motif per company standard procedures, and the sequencing raw data satisfactorily met quality control. Chromatin was isolated after adding lysis buffer, followed by disruption with a Dounce homogenizer. Genomic DNA was sheared to an average length of 300–500 bp by sonicating the lysates, and the segments of interest were immunoprecipitated using an antibody (4 μg) against BRD4, H3K27ac, H3K27me3, or H3K4me1. The protein/DNA complexes eluted from beads were treated with RNase and proteinase K, crosslink was reversed, and the ChIP DNA was then purified for use in the preparation of Illumina sequencing libraries. Standard steps included end-polishing, dA-addition, adaptor ligation, and PCR amplification.