Western Blot Analysis

Western blot analysis using specific antibodies against Akt, Akt1, Akt2, GSK-3β, and GS, phosphorylated (p-)Akt (Ser⁴⁷³), -Akt1 (Ser⁴⁷³), -Akt2 (Ser474), –GSK-3β, and -GS, hexokinase II (HK II; Cell Signaling Technology, Danvers, MA), TRB3 (EMD Millipore, Billerica, MA), connective tissue growth factor (CTGF), transforming growth factor-β1 (TGF-β1), plasminogen activator inhibitor-1 (PAI-1), tumor necrosis factor-α (TNF-α), peroxisome proliferator–activated receptor γ coactivator-1α (PGC-1α), GAPDH, β-actin (Santa Cruz Biotechnology, Santa Cruz, CA), and MT (DakoCytomation, Santa Clara, CA) was performed as described previously (14–16,18,19).