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. 2017 Jun 26;8:15926. doi: 10.1038/ncomms15926

Figure 3. PPARγ transcriptionally controls WAT niche expansion and APC niche interaction.

Figure 3

(a) Microarray heat map of AT-GFP+ (adipose progenitor) cells isolated from 2-month-old AT-control and AT-PPARγ-LOF mice (n=6 mice per group in triplicate). (b) GFP− and GFP+ cells were FACS isolated from AT-GFP control mice and adipocytes were isolated by floatation. PDGFRβ mRNA expression was measured. (c) GFP+ cells were FACS isolated from AT-Control, Sox-PPARγ-LOF, AT-PPARγ-LOF, SMA-PPARγ-LOF and AT-PPARγ-GOF and AT-PPARγ-Rescue or SV cells were isolated from SMA-PPARγ-GOF and PDGFRβ mRNA expression was assessed. (d) Control, Sox-PPARγ-LOF, AT-PPARγ-LOF, AT-PPARγ-GOF and AT-PPARγ-Rescue (n=10 per group) were administered normal chow or rosiglitazone (0.0075% diet) for 7 days. Subsequently, GFP+ cells were FACS isolated and PDGFRβ mRNA expression was measured. (e) GFP+ cells were FACS isolated from AT-GFP control mice (n=8) and treated with the denoted concentrations of rosiglitazone for 4 h. mRNA expression of denoted genes were measured. (f) GFP+ cells isolated from AT-control mice (n=8) 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. (g) GFP+ cells were FACS isolated from AT-control mice and cultured. 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. (h) Subcutaneous IGW explants were excised from AT-control mice and encased in Matrigel. Explants were treated with vehicle (5% DMSO), PDGF-B (10 ng ml−1), SU16F (5 μM) or both for 5 days and vascular sprouting was assessed. Scale bar 100 μm. Data are expressed as means±s.e.m. *P<0.05 unpaired t-test, two tailed: GFP+ and adipocytes compared to GFP− SV cells. **P<0.05 unpaired t-test, two tailed: mutants compared to AT-control mice. #P<0.01 unpaired t-test, two tailed: rosiglitazone treated compared to chow treated. §P<0.01 unpaired t-test, two tailed: vehicle compared to IgG control.