Fig 1: Effect of GW501516 on BCAA transport and catabolic enzymes. (a and b) Effect of treatment with GW501516 (GW) at 1 μM for 24 hours on (a) absolute media BCAA content or (b) control mean-normalized (within each experiment) media BCAA content following 24-hour treatment. (c) Effect of GW at 1 μM for up to 24 hours on myotube mRNA expression of branched-chain aminotransferase 2 (Bcat2), branched-chain alpha-keto acid dehydrogenase (Bckdha), and 3-hydroxyisobutyrate dehydrogenase (Hibadh). (d) Effect of GW at 1 μM for 24 hours on myotube protein expression of large amino acid transporter 1 (LAT1), pBCKDHa (normalized to total BCKDHa), BCKDHa, and BCAT2. Notes: ∗ indicates p ≤ 0.05 between groups. Time course gene expression was analyzed using one-way ANOVA with Dunnett's correction for multiple comparisons. Target gene expression was normalized to tata binding protein (Tbp) using three replicates per group across two independent experiments with n = 5–6 for the final analysis. Protein expression and BCAA media content were analyzed using student's t-test. Western blots were performed using three replicates per group across two independent experiments with n = 6 for the final analysis. BCAA media content was performed using three replicates per group across two independent experiments with n = 6 for the final analysis with each analyte measured in triplicate.