Anti srebf1

To obtain total protein lysates, triturated tissue or cell pellets were lysed with cell lysate‐containing mixed protease inhibitors. The lysate was incubated on ice for 30 min and centrifuged at 12,000 g for 15 min at 4°C. The protein concentration of each sample was determined using the bicinchoninic acid (BCA) assay. Samples were loaded on 10% SDS‐PAGE. After electrophoresis, proteins in the gel were transferred to PVDF membranes. Membranes were blocked with 5% skimmed milk in TBST for 2 h at room temperature, then incubated with primary antibodies overnight at 4°C, followed by secondary antibodies for 2 h. The main antibodies include anti‐ACLY (Proteintech, Wuhan, China), anti‐SREBF1 (Proteintech, Wuhan, China), anti‐MOGAT2 (Proteintech, Wuhan, China), anti‐ACC1 (Proteintech, Wuhan, China), anti‐ACS (Proteintech, Wuhan, China), anti‐FASN (Proteintech, Wuhan, China), anti‐β‐actin (Proteintech, Wuhan, China), pan‐acetylated antibody (AC) (PTMBIO, Hangzhou, China), anti‐IgG (Proteintech, Wuhan, China), anti‐SIRT2 (Proteintech, Wuhan, China), anti‐HDAC1 (Proteintech, Wuhan, China) and anti‐PCAF (Proteintech, Wuhan, China). Immunoreactive bands were observed with an Azure C300 system.

The following antibodies and reagents were used: Anti-SREBF1 (Proteintech, 14088-1-AP, 1:1000 for western blotting and 4 μg for ChIP), anti-KLF5 (Santa Cruz Biotechnology, sc-398409X, 1:1000 for western blotting, and 4 μg for ChIP), anti-TP63 (R&D Systems, AF1916-SP, 1:1000), anti-Actin (Santa Cruz Biotechnology, sc-8432, 1:2000), anti-ACLY (Cell Signaling Technology, 4332, 1:1000), Anti-FASN (Cell Signaling Technology, 3180, 1:1000), anti-GAPDH (Cell Signaling Technology, 2118, 1:2000), anti-mTOR (Cell Signaling Technology, 2972S, 1:1000), anti-Phospho-mTOR (S2448) (Cell Signaling Technology, 5536T, 1:1000), anti-Phospho-MEK1 (Ser298) (Cell Signaling Technology, 9128S, 1:1000), anti-MEK1/2(D1A5) Rabbit (Cell Signaling Technology, 9124S, 1:1000), anti-p70 S6 Kinase (Cell Signaling Technology, 9202S, 1:1000), anti-Phospho-p70 S6 Kinase (Cell Signaling Technology, 9205S, 1:1000), anti-mouse IgG-HRP (Jackson ImmunoResearch Laboratories, Inc., 115-035-003, 1:10000), anti-rabbit IgG-HRP (Jackson ImmunoResearch Laboratories, Inc., 111-035-144, 1:10000), anti-goat IgG-HRP (Jackson ImmunoResearch Laboratories, Inc., 705-035-003, 1:10000), HCS LipidTOX™ Green Neutral Lipid Stain (Thermo Scientific, H34475, 1:100), Lipofectamine RNAiMAX (Thermo Scientific, 13778150), and Fatostatin (Cayman Chemical, 12562).