Figure 1 | Cell Discovery

Figure 1

From: Retinoic acid induces white adipose tissue browning by increasing adipose vascularity and inducing beige adipogenesis of PDGFRα+ adipose progenitors

Figure 1

RA promotes in vitro angiogenesis of SVCs via VEGFA/VEGFR2 signaling. (a) VEGFA protein content in the medium cultured with adipose-derived SVCs for 2 days, and cells were treated with different doses of RA. (b) Quantification of VEGFA protein, data presented were normalized according to the content of a non-specific band. (c) The expression of Vegfa in SVCs treated with RA for 2 days. (d) Representative images (scale bar=100 μm), and (e) Vegfa expression of adipose tissue derived control or Vegfr2 knockout SVCs cultured on matrigel-coated plates in endothelial basal medium for 2 days with or without 1 μm RA (same doses for other in vitro studies if unstated). (f) Putative retinoic acid response elements (pRARE) on the Vegfa promoter are indicated by letters, some of which share several nucleotides; closely located pRARE were analyzed using one pair of primers. (g) SVCs were treated with RA or DMSO for 4 h, and binding of RAR and retinoid X receptor (RXR) to pRAREs were analyzed by ChIP. (h) The pRARE sites with altered RAR/RXR binding due to RA were chosen. These sites on a Vegfa promoter luciferase reporter plasmid were mutated. The luciferase activity of WT and mutated Vegfa promoter reporter plasmids were analyzed after 4 h of RA treatment. Data presented are mean±s.e.m., n=6, *P<0.05.

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