Comprehensive Western Blot Analysis of Key Metabolic Regulators

Western blot analysis was performed as described previously^{8 (link)}. Proteins were separated by SDS-PAGE, transferred to a PVDF membrane (Millipore, Billerica, MA, USA), and incubated with specific primary antibodies [anti-SREBP-1c (577036, BD Biosciences, Franklin Lakes, NJ, USA), anti-FAS (3180, Cell Signaling, Danvers, MA, USA), anti-C/EBPα (2295, Cell Signaling), anti-PPARγ (2435, Cell Signaling), anti-PDK1 (KAP-PK112, Enzo Life Science, Farmingdale, NY, USA), anti-PDK2 (sc-100534, Santa Cruz, Dallas, TX, USA), anti-PDK3 (sc-365378, Santa Cruz) and anti-PDK4 (ab214938, Abcam), anti-p-PDHE1α Ser232 (AP1063, Calbiochem, San Diego, CA, USA), anti-p-PDHE1α Ser293 (AP1062, Calbiochem), anti-HIF1α (NB100-123, Novus Biologicals, Centennial, CO, USA), anti-HSP90 (4874, Cell Signaling), and anti-β-tubulin (G098, Applied Biological Materials, Richmond, BC, Canada)].