Figure 3

Fasting induces adipose-VEGF expression. (A) MA plot highlighting significantly altered mRNA expression of adipose-derived factors in PWAT of HFD-IF mice, compared to that of HFD-AL mice. Vegfa, Cfd, Nrg4, Adipoq, and Lep encode vascular endothelial growth factor, adipsin, neuregulin 4, adiponectin, and leptin, respectively. (B) qPCR validation of adipose-derived factors in PWAT (HFD, AL/IF: n = 6/8). (C) mRNA expression levels of Vegfa, Cfd, Nrg4, Adipoq, and Lep in PWAT at feeding and 24 h of fasting (n = 5 per group). (D) Vegfa mRNA expression in PWAT at different fasting durations (n = 5-6 per group). (E) Vegfa mRNA expression in PWAT at feeding, 24 h of fasting, 6 h of refeeding, fasting with β3-AR antagonist, SR59230A (5 mg/kg, i.p.), and fasting with non-specific β-AR antagonist, Propranolol (5 mg/kg, i.p.; n = 5 per group). (F) Vegfa mRNA expression in PWAT at feeding and 24 h of fasting with treatments of vehicle or clodronate (n = 5). (G) A representative macroscopic image illustrating increased vascularization in IWAT of HFD-IF mice, compared to HFD-AL mice. Black and white arrows indicate IWAT of HFD-AL and HFD-IF mice, respectively. (H) Representative microscopic images of adipocytes and blood vessels, visualized with perilipin and PECAM-1 antibodies, respectively, in whole-mount PWAT. (I) Quantification of vessel densities in PWAT. Data are mean ± SEM; one- or two-way ANOVA with Student-Newman-Keuls post hoc analysis and two-tailed unpaired Student's t-test; ^*P < 0.05 vs HFD-AL or Fed. ^#P < 0.05 vs Fast (24 h).