GSM9791793: D-galactose plus MCHA co-treated L929 fibroblast, NFIX immunoprecipitation ChIP sample; Mus musculus; ChIP-Seq
L929 cells from Control, D-gal, and D-gal+MCHA groups were cross-linked with 1% formaldehyde for 10 min at room temperature, followed by quenching with glycine. Chromatin was isolated and sheared to an average size of 200–500 bp using a sonicator. Immunoprecipitation was performed overnight at 4°C using a specific antibody against NFIX and Protein A/G magnetic beads. For Input controls, 10% of the sheared chromatin was saved before immunoprecipitation. Cross-linking was reversed, and DNA was purified using a PCR purification kit. Sequencing libraries were generated using the NEBNext Ultra II DNA Library Prep Kit for Illumina] following the manufacturer's instructions. Briefly, purified ChIP DNA and Input DNA underwent end-repair, 3' adenylation, and ligation with Illumina-compatible adaptors. The adaptor-ligated fragments were then enriched by limited-cycle PCR amplification. Library quality and size distribution were assessed using an Agilent 2100 Bioanalyzer, and quantified by qPCR.