Anti srebf1

Chromatin immunoprecipitation (ChIP) assays were performed by using the ChIP assay kit (Beyotime, Shanghai, China) according to the manufacturer's protocols (http://www.beyotime.com/product/P2078. htm). Briefly, GMEC were seeded onto 100 mm × 20 mm cell culture dishes and cultured until 90 to 100% confluence. Then, cells were fixed with 1% formaldehyde for 10 min at 37°C. The cells were washed with cold PBS and cell lysates were sonicated to produce chromatin fragments averaging 200 to 1,000 bp. The fragmented chromatin was then added to ChIP dilution buffer (Beyotime) and the samples were incubated overnight at 4°C with anti-SREBF1 (Abcam, Cambridge, MA) and normal mouse IgG (Millipore, Darmstadt, Germany) antibodies. The resultant immune complexes were precipitated with protein A+G agarose/Salmon Sperm DNA (Beyotime) and washed with wash buffer (Beyotime). Finally, the DNA-protein crosslinks were reversed by incubating the samples in 5 mM NaCl at 65°C for 4 h. The DNA was then treated with proteinase K (Millipore) at 45°C for 1 h and precipitated chromatin was used as the template for PCR with the primers listed in Table 2.