SREBP1 ChIP-seq in HEK293 cells

Human Flag-SREBP1a CDS plasmid was transfected into HEK293T cells for 48h and ChIP assays were performed with the ChIP kit. ChIP-enriched DNA samples were used to construct sequencing libraries with the NEBNext UltraTM DNA Library Prep Kit for Illumina. Briefly, NEBNext Adaptor and Ligase Master Mix were added to ligate adaptors, and USER Enzyme was also added to remove uracil bases in the adaptors. Libraries were purified with AMPure XP magnetic beads, followed by 10 cycles of PCR amplification, depending on input DNA amount. Amplified libraries were size-selected and purified using AMPure XP Beads. Library quality and concentration were assessed using Qubit and High-Sensitivity DNA chips. Paired-end 150 bp (PE150) sequencing was performed on the Illumina NovaSeq X Plus sequencing instrument. Raw reads from IP and input samples were quality-controlled to remove adapters, repetitive sequences, and low-quality reads, to obtain clean data. Subsequently, clean reads were aligned to the reference genome using Bowtie 2 (version 2.2.0), generating BAM alignment files. Duplicate reads were removed, and MACS2 (version 2.2.8) was used for peak calling.