Sc 23016

Western blot analysis was conducted essentially as described (Xu et al., 2015) . Briefly, protein was isolated from liver tissue homogenized in RIPA lysis buffer (Beyotime, Shanghai, China). Equal amounts of protein were separated on 10% SDS polyacrylamide gels to resolve SREBP1 and SCD1. Proteins were transferred onto nitrocellulose membranes (Millipore, Billerica, MA), which were incubated with primary antibodies against either SCD1 (Goat polyclonal antibody, sc-23016, Santa Cruz Biotechnology, Santa Cruz, CA; diluted to 1:200) or SREBP1 (Mouse monoclonal antibody, ab3259, Abcam, Cambridge, UK; diluted to 1:1,000). After extensive washing, blots were incubated with horseradish peroxidase-coupled secondary antibodies. We used affinity-purified rabbit anti-goat IgG antibodies (E030130-01, Earth Ox, San Francisco, CA; diluted to 1:10,000) to detect SCD1 and rabbit antimouse antibodies (ab6728, Abcam; diluted to 1:5000) to detect SREBP1. Differences in protein transfer efficiency between blots were normalized based on quantification of GAPDH, which was detected with rabbit polyclonal antibodies (AP0066, Bioworld, St. Louis Park, MN; diluted to 1:10,000). Intensities of bands from target genes were quantified with the Quantity One 1-D analysis software (Bio-Rad, Hercules, CA).