Figure 2. Blocking MAT expansion directly prevents increased circulating adiponectin during CR.

WT and Ocn-Wnt10b mice were fed ad libitum or a 30% CR diet from 9-15 weeks of age. (A) Masses of iWAT, gWAT and BAT in 15-week-old mice. (B,C) BM adiposity was assessed by osmium tetroxide staining of tibiae followed by μCT analysis. (B) Representative images of stained tibiae; scale bar = 1 mm. (C) MAT as % marrow volume (MV) was determined from μCT scans. (D) Analysis of HMW, MMW, LMW and total adiponectin in sera of 15-week-old mice. Immunoblots are from the same exposure of film. (E) Quantitation of serum adiponectin from (D). (F,G) qPCR analysis of Adipoq expression in iWAT, gWAT, or combined tibiae and femurs (Tib/Fem). (H-J) Total RNA and protein was isolated from quadriceps muscle. Expression of Pgc1a, Tfam and Acadm was determined by qPCR (H). Protein phosphorylation was determined by immunoblotting (I) and quantified by densitometry (J). CaMKII phosphorylation is a marker of Ca²⁺/calmodulin signaling. Data are mean ±SEM of the following numbers of mice: WT, n = 6; Wnt10b, n = 4; WT CR, n = 5; Wnt10b CR, n = 6. Similar results were observed in a second mouse cohort. For each diet group, significant differences between WT and Wnt10b mice are indicated by * (P <0.05), ** (P <0.01) or *** (P <0.001). Within each genotype, significant differences between ad libitum and CR diets are indicated by ## (P <0.01) or ### (P <0.001). See also Figure S2, Figure S3, and Table S1.