Figure 5. Microarray and effect of AdVEGF delivery on adipose angiogenesis and CL-induced PDGF-C expression during beige transition.

(a) Histological images of microvessels (CD31⁺ red, white arrows and arrow heads), adipocyte morphology (H&E, double-arrowed bars, perilipin+green), and UCP1 (UCP1⁺ red, white arrows) in mice that were treated with AdGFP and AdVEGF for 10 days. The non-adenovirus-treated (NAT) group served as a control. n=5 mice for NAT group, n=10 mice for each AdGFP and AdVEGF group. (b) Quantification of Vegf mRNA expression levels in NAT- AdGFP- and AdVEGF-treated total gWAT. Quantification of microvessel density in NAT- AdGFP- and AdVEGF-treated (n=10 random fields; n=5 mice for NAT and n=10 for each AdGFP and AdVEGF group). (c) Quantification of UCP1-positive signals (n=10 random field; n=5 mice for NAT group and n=10 for each AdGFP and AdVEGF group) and quantification of Ucp1 mRNA expression levels by qPCR (n=10 samples for each group). (d) Quantification of average adipocyte size (>30 adipocytes/field; n=10 random fields; n=5 mice for NAT group and n=10 for each AdGFP and AdVEGF group). (e) Norepinephrine-stimulated non-shivering thermogenesis in AdGFP or AdVEGF-treated gWAT of mice (n=5 mice for each group). NE, norepinephrine. (f) Hierarchical clusters of top 20 growth factors and cytokines from genome-wide microarray analysis (n=3 samples for each group). qPCR analysis of Pdgf-c mRNA expression levels of vehicle- and CL-treated gWAT SVF (n=5 samples for each group). (g) Quantification of Pdgf-c mRNA expression levels in SVF by qPCR in AdGFP- and AdVEGF-treated total gWAT (n=10 samples). (h) QPCR analysis of Pdgf-a, -b, -c, and -d expression levels in total vehicle- and CL-treated CD31⁺ EC fraction from gWAT. (i) Pdgf-c mRNA expression levels in vehicle- and CL-treated total gWAT of wt and Flk1f/f;Tie2CreER^T2 mice. All scale bars, 50 μm. *P<0.05; **P<0.01 by two-sided unpaired t-test. Data presented as mean±s.e.m. n.s., not significant.