Anti pparα

Total protein was extracted from liver tissues or HepG2 cells. Samples were separated using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis and electroblotted onto polyvinylidene difluoride membranes. The membranes were first blocked with non‐fat milk for 2 h, and incubated with the following primary antibodies: anti‐PPARγ, anti‐PPARα, anti‐CD36(ABclonal Technology, Boston, MA, USA), anti‐GAPDH and anti‐α‐tubulin (Beijing Ray antibody, Beijing, China), followed by incubation with a horseradish peroxidase‐conjugated anti‐rabbit or mice secondary antibody (Beijing Ray antibody). Objective proteins were detected using ECL Plus (Pierce, NJ, USA).

Liver tissue were lysed in RIPA buffer containing a protease inhibitor (Beyotime Institute of Biotechnology, China). 30 μg of total protein were separated using a sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and followed by the transfer of proteins to a PVDF membrane. After blocking with 5% skim milk, membranes were incubated with the appropriate primary antibodies overnight at 4 °C. Primary antibodies including anti-phospho-IRS-1^Ser612(CST), anti-IRS-1(CST), anti-phospho-AKT^Ser473(CST), anti-AKT (CST), anti-Glut-2 (Abclonal), anti-Sirt1(Abcam), anti-PPARα (Abclonal), anti-CPT1α(Abcam), anti-AMPK (Abcam). Blots were incubated with an appropriate horseradish conjugated secondary antibody (Beyotime Institute of Biotechnology, China) at room temperature for 1 h. Antibody bound protein was detected by ECL (Merkmillipore) and quantified by scanning densitometry. Relative protein expression was normalized to Tublin expression.

Primary antibodies used were anti–Lamin A/C (catalog 2032), rabbit monoclonal anti–p-p38 MAPK (Thr180/Tyr182) (catalog 9215), anti–p-SAPK/JNK (Thr183/Tyr185) (catalog 9255) (all Cell Signaling Technology); anti–β-actin (catalog A5441) and anti–p-PPARγ S112 (catalog 04-816-I) (both Sigma-Aldrich); anti–p-ERK (sc-7383, Santa Cruz Biotechnology); anti-PPARγ (A0270, Abclonal); and anti-PPARα (catalog PAa-822A), anti-PPARδ (PA1-823A), and anti–p-PPARγ S273 (catalog BS-4888R) (all Thermo Fisher Scientific). See complete unedited blots in the supplemental material.