FIGURE 3.

PPARγ overexpression partially rescues the loss of adipogenesis with chemerin or CMKLR1 knockdown. Prior to initiating adipogenesis, BMSCs were treated with the adenoviral transduction VEH (Polybrene; 6 μg/ml) or transduced with GFP- or PPARγ-overexpressing adenovirus concomitant with knockdown of LacZ (LZ), chemerin (CHE), or CMKLR1 (CMK) expression using adenovirally delivered shRNAs. Oil red O phase-contrast images (A) and quantitation of this staining (B) revealed the inability of PPARγ to fully rescue adipogenesis in cells lacking chemerin or CMKLR1 expression. Chemerin, adiponectin, and PPARγ mRNA levels were quantified after a 14-day differentiation period using qPCR and are expressed relative to VEH control (C–E). Chemerin and adiponectin protein levels were measured by immunoblot of equal volumes of the adipocyte medium after 14 days of differentiation (F). Total lysates of the same cells were used to quantify PPARγ and β-actin (loading control) protein levels by immunoblot (F). Error bars represent S.E. (n = 4). *, p < 0.05 versus respective GFP control.