Goat anti mouse and goat anti rabbit hrp conjugated secondary antibodies

Proteins were extracted from the liver tissue of the mice [12 ,13 (link)]. The livers were homogenized in RIPA buffer, 10 mM of NaF, 1 mM of Na₃VO₄, 1 mM of PMSF, and protease inhibitor tablets (Roche Diagnostics). The protein concentration was determined using a BCA protein assay kit (Thermo Fisher Scientific), and lysates were analyzed using SDS-polyacrylamide gel electrophoresis and Western blotting analysis on PVDF membranes. Antibody for PKCβ (F-7) was purchased from Santa Cruz Biotechnology, and antibodies for AKT (#4685), P-AKT^Thr308 (#13038), P-AKT^Ser473 (#4060), insulin receptor beta (#3025), P-insulin receptor/IGF1R beta (#3021), P-IRS-1^Ser307 (#2381), P-IRS-1^Ser612 (#3203), P-IRS1^Ser318 (#5610), IRS-2 (#4502), P-mTOR^Ser2448 (#5536), P-mTOR^Ser2481 (#2974), mTOR (#2983), rictor (#2114), and GβL (#3274) were purchased from Cell Signaling Technology (Danvers, MA, USA). Phospho-SGK1^Ser422 (#55281) and SGK1 (#43606) were purchased from Abcam (Cambridge, MA, USA). Goat anti-mouse and goat anti-rabbit HRP-conjugated secondary antibodies (Bio-Rad) were used.

Proteins were extracted from liver tissue of mice (16 (link), 17 (link), 18 (link), 19 ). Livers were homogenized in RIPA buffer, 10 mM NaF, 1 mM Na₃VO₄, 1 mM PMSF, and protease inhibitor tablet (Roche Diagnostic). Protein concentration was determined using a BCA protein assay kit (Thermo Scientific), and lysates were analyzed by SDS-polyacrylamide gel electrophoresis and Western blot analysis on a PVDF membrane. Antibody to UCP-1 (14670S), UCP-2 (89326S), UCP-3 (970000S), P-ERK-1/2^{Thr202/Tyr204} (9101), and ERK-1/2 (#9102) were purchased from Cell Signaling Technology. Antibody to PKCβ (ab38279) and FGF21 (ab1711941) were purchased from Abcam, Cambridge, MA, and antibody to PGC-1α (A11971) was purchased from ABclonal. Goat anti-mouse and goat anti-rabbit HRP-conjugated secondary antibodies (Bio-Rad) were used.