Anti acadl

Whole protein lysates from the mouse liver, serum, and Hepa-1c1c7 cells were extracted using lysis buffer supplemented with protease and phosphatase inhibitors (Sigma-Aldrich). Aliquots containing 30 μg of proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transblotted onto polyvinylidene difluoride membranes (Bio-Rad, Hercules, CA), blocked with 4% nonfat dry milk in phosphate-buffered saline solution containing 1% Tween-20, and then incubated overnight with the following antibodies, including anti-DGAT1, anti-ApoB100 (Santa Cruz Biotechnology, Dallas, TX), anti-ACSL1, anti-LAMP1, anti-LAMP2, anti-ATF4, anti-CHOP (Cell Signaling Technology, Danvers, MA), anti-ACADL, anti-ACOX1 (Proteintech Group, Inc, Rosemont, IL), anti-ACADM, anti-LC3II (Novus Biologicals, Littleton, CO), anti-β-actin, and anti-GAPDH (Abcam, Cambridge, MA), respectively. Membranes were then washed and incubated with horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G or goat anti-rabbit immunoglobulin G (Thermo Fisher Scientific, Rockford, IL). The bound complexes were detected with enhanced chemiluminescence (Thermo Fisher Scientific) and quantified by densitometry analysis.