Anti fasn antibody

Cells were lysed using radioimmunoprecipitation buffer (Beyotime, Shanghai, China). Total protein (20 μg) was separated by 8% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred onto 0.45 μm polyvinylidene fluoride membranes (Millipore, Billerica, USA) using a Mini-Protean System (Bio-Rad, Hercules, USA). Membranes were incubated with primary antibody (1:500 dilution, catalog no. ABS1508, anti-SREBP1 antibody from Sigma-Aldrich; 1:500 dilution, catalog no. 3189S, anti-FASN antibody from Cell Signaling, Danvers, USA; 1:1 200 dilution, catalog no. AA128, anti-ACTB antibody from Beyotime) overnight at 4°C, then incubated with horseradish peroxidase-conjugated anti-rabbit antibody from Beyotime (1:8 000 dilution, catalog no. A0208) or anti-mouse antibody from Beyotime (1:12,000 dilution, catalog no. A0216) for 2 h at 37°C. Enhanced chemiluminescence (Beyotime) was added to the membranes, which were detected using an Odyssey Fc Imaging System (Li-Cor, Lincoln, USA) [22 (link)].

WB was performed as previously described [29 (link)]. Each experiment was successfully carried out three times, as indicated. The following antibodies were used for WB: anti-HMGCS2 antibody (1:1000, Abcam, Cambridge, MA, USA), anti-ACC antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-phospho (S79) ACC antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-FASN antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-Lipin1 antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-SREBP2 antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-ACSL1 antibody (1:1000, Cell Signaling, Beverly, MA, USA), anti-GAPDH antibody (1:2000, Proteintech, Chicago, IL, USA) anti-α-tubulin antibody (1:5000, Sigma-Aldrich, St. Louis, MO, USA). The ImageJ software (National Institutes of Health, Bethesda, MD, USA) was used to quantify the images.