Figure 7: VEGF is a transcriptional target of PPARγ in APCs.

(a) GFP− and GFP+ cells were FACS isolated from AT-GFP control mice and adipocytes were isolated by floatation (n=6). VEGF mRNA expression was measured. (b) AT-GFP+ cells were FACS isolated from AT-GFP control mice (n=6) and treated with vehicle or VEGF for 8 h and then treated with BrdU for 12 h. BrdU incorporation was monitored. (c) AT-GFP+ cells were FACS isolated from AT-GFP control mice (n=6) and treated with vehicle or VEGF for 8 h and then plated in transwell migration chambers. Migration was monitored 12 h later and quantified. (d–f) Total SV cells were isolated from AT-GFP control mice (n=6). Cells were treated with vehicle or VEGF for 12 h and then mRNA was harvested or cells were imaged for AT-GFP and stained for CD31. (g) Cells and staining described in d were quantified for AT-GFP and CD31 co-localization. (h) AT-GFP+ cells were FACS isolated from AT-control, SMA-PPARγ-LOF and AT-PPARγ-GOF and AT-PPARγ-Rescue or SV cells were isolated from SMA-PPARγ-GOF and VEGF mRNA expression was assessed (n=6 per group). (i) AT-GFP+ cells were FACS isolated from AT-control mice (n=6) and treated with denoted concentrations of rosiglitazone for 4 h. mRNA expression of VEGF and Angptl4 were measured. (j) AT-GFP+ cells isolated from AT-control mice (n=6) and were pretreated with cyclohexamide (10 μg ml⁻¹) for 15 min and then treated with 1 μM rosiglitazone for 4 h and PDGFRβ mRNA expression was measured. (k) AT-GFP+ cells were FACS isolated from AT-control mice (n=6). Cells were then treated with vehicle (dimethylsulfoxide (DMSO)) or 1 μM rosiglitazone for 4 h and then ChIP-qPCR analysis was performed to assess PPARγ occupancy. Data are expressed as means±s.e.m. Scale bar 100 μm. *P<0.05 unpaired t-test, two tailed: GFP+ and adipocytes compared to GFP− SV cells. P<0.01 unpaired t-test, two tailed: VEGF treated compared to vehicle treated cells. **P<0.02 unpaired t-test, two tailed: mutant mice compared to control mice. §P<0.001 unpaired t-test, two tailed: vehicle compared to IgG control. #P<0.001 rosiglitazone treated compared to vehicle (DMSO) treated.