Glut2 07 1402 and pex14 abc 142

Protein lysates were prepared from frozen powdered liver using T-PER buffer (ThermoFisher 78510) containing protease inhibitor cocktail and phosphatase inhibitors from Sigma-Aldrich (St. Louis, MO). Western blot analyses were performed using standard PAGE-SDS as previously described (25 (link)). Primary antibodies were as follows: ACC (3676), p-ACC (11818), AMPKα (5831), p-AMPKα (2535), AMPKβ1/2 (4150), p-AMPKβ1 (4181), and Pex5 (83020) from Cell Signaling; Catalase (ab16731), total OXPHOS cocktail (ab110413), Pex19 (ab137072), and PMP70 (ab3421) from Abcam; GLUT2 (07-1402) and Pex14 (ABC-142) from EMD Millipore, and PGC-1α (sc-517380) from Santa Cruz Biotechnology. Horseradish peroxidase-linked secondary antibodies (anti-mouse IgG Cat NXA931 and anti-rabbit IgG Cat NA934V) were purchased from GE Healthcare (Piscataway, NJ) and proteins were detected using enhanced chemiluminescence chemistry. Reversible protein stain-MemCode (MemC) (Thermo-Fisher 24580) was used to confirm equal transfer of proteins and quantitation of these bands served as a loading control. A ChemiDoc imaging system (Bio-Rad) was used to image bands and Image Lab software (Bio-Rad) was used to quantitate band intensity. Data are expressed as fold change compared with the Con-Sed group after normalizing to MemC.