FIGURE 2.

A2bAR agonism inhibits adipogenesis of SVCs. SVCs were isolated from the subcutaneous adipose tissue of WT mice, treated with vehicle (DMSO), 1 μm BAY 60-6583 (BAY), or 10 μm CGS 21680 (CGS) on day 0 and day 3 of induction, and induced to differentiate as described under “Experimental Procedures.” A, representative image of Oil Red O staining performed 6 days after induction of differentiation. B–F, mRNA expression of PPARγ (B), C/EBP-α, (C), aP2 (D), adiponectin (E), and perilipin (F) after 6 days of induction. Agonist treatment was determined using the ΔΔCT method and normalized to 18 S rRNA values (n = 3). G, representative Western blot analysis for PPARγ (57 kDa), aP2 (15 kDa), and C/EBP-α (45 kDa) performed with β-actin (45 kDa) as a loading control. (The blot was stripped and reprobed twice, including with anti β-actin.) H–J, quantification of Western blot analysis results was performed with ImageJ software and normalized to β-actin (n = 4). AU, arbitrary units. K, mRNA expression of aP2 after 6 days of induction and agonist treatment was determined using the ΔΔCT method and normalized to 18 S rRNA values (n = 4). Data are mean ± S.E. *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared with vehicle. Analyses were performed by Student's t test (B—F and H–J) or by one-way ANOVA with Bonferroni multiple comparisons post hoc test (K).