PPARβ/δ Agonism with GW501516 Increases Myotube PGC-1α Content and Reduces BCAA Media Content Independent of Changes in BCAA Catabolic Enzyme Expression

Figure 9

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 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 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 for the final analysis. BCAA media content was performed using three replicates per group across two independent experiments with for the final analysis with each analyte measured in triplicate.

(a)

(b)

(c)

(d)