GSM9229005: 3T3-L1,  Sirt3-OE 2 Input; Mus musculus; ChIP-Seq
A ChIP assay was conducted utilizing a kit from Cell Signaling Technology, USA (catalog number 91820S). 3T3-L1 adipocytes were fixed using 4% formaldehyde and subsequently harvested by scraping for lysis. Cross-linked chromatin samples were extracted using nuclease digestion (37 °C for 20 min) and sonication (20% power for 5 s, followed by 10 s, repeated 30 times in total). Soluble fragmented soluble chromatin was immunoprecipitated for 48 h at 4 °C with H3K56ac antibody and incubated with Protein A/G for 2 h. The beads were then resuspended in ChIP elution buffer, and cross-link reversal was performed at 65 °C for 30 min. The enriched DNA was isolated by incubation with proteinase K and RNase A. After end-repair and A-tailing, the DNA was treated with sequencing adapters and amplified by PCR to obtain the sequencing library. The library was quantified with the help of Qubit2.0, and the MGISEQ-T7 sequencer was used to sequence the various libraries in accordance with the effective concentration as well as the target data volume. After aligning the reads to the reference mouse genome, the ChIP-seq peaks were identified using MACS version 1.4.2.