Fig. 7.

VEGF signaling plays an important role in RA regulation of adipogenesis. (a) Representative PDGFRα and VEGFR2 immunohistochemical staining images of E12.5 embryo treated with MVA. (b) FACS analysis of PDGFRα ⁺/VEGFR2⁺ cells (CD45⁺ cells were excluded) in iWAT of maternal RA or BMS493 treated (at E10.5) fetuses at E12.5 (numbers in the circles indicate the percentages of cells). (c) FACS analysis of PDGFRα⁺/VEGFR2⁺ cells in iWAT of weanling MVA offspring. (d) VEGFR2 was knocked out in P19 cells, then embryo bodies were formed and pretreated with RA for 3 days and, then, induced brown adipogenesis for 4 days. Prdm16 expression was analyzed by qRT-PCR (n = 3). (e–g) Vegfr2 were knocked out in PDGFRα⁺ cells at E10.5 by an injection of 20 mg/kg BW tamoxifen to the pregnant mothers. Adipose tissue weight was measured at weaning (e, n = 6). Brown adipose gene expression in iWAT was analyzed by qRT-PCR (f, n = 6). Adipose tissue sections were stained by H&E or IHC using anti-UCP1 antibody (g). Data presented are mean ± SEM, unpaired two-tail t-test, *p < 0.05.