GSM9420688: OB_TNFa_p65_1; Mus musculus; ChIP-Seq
Cells were crosslinked with formaldehyde and ChIP was performed using the SimpleChIP® Plus Enzymatic Chromatin IP Kit with magnetic beads (Cell Signaling Technology) according to the manufacturer's protocol. Equal amounts of chromatin (equivalent to chromatin from ~4 × 10⁶ cells) were immunoprecipitated using ChIP-validated anti-Smad4 (D3R4N) and anti-p65 (D14E12) antibodies. After reverse crosslinking and proteinase K treatment, DNA was purified using ChIP DNA Clean & Concentrator (Zymo Research, Irvine, CA, USA). Library preparation, sequencing, and initial processing were performed by Novogene. Libraries were sequenced as 150-bp paired-end reads on a NovaSeq platform to a depth of >10 million mapped reads per sample.