Figure 6: Co-localization of NFIA facilitates PPARγ binding to the brown-fat-specific enhancers and drives active transcription.
From: NFIA co-localizes with PPARγ and transcriptionally controls the brown fat gene program
(a) C2C12 myoblasts expressing only PPARγ —and those expressing both PPARγ and 3×FLAG-NFIA—were stained with Oil Red O seven days after inducing adipocyte differentiation. Scale bar, 50 μm. (b) Indicated genes were quantified by RT-qPCR at day 7 after adipocyte differentiation (mean ± s.e.m.; n = 3 independent samples; ∗∗P < 0.01; NS, not significant). (c) Western blot analysis of the PPARγ protein in PPARγ − or PPARγ + 3×FLAG-NFIA-expressing C2C12 myoblasts. β-actin was used as a loading control. (d) ChIP-qPCR analysis of NFIA. Cidea 29 kb, Ppara 21 kb, Ppargc1a − 97 kb and Ucp1 9.5 kb are background sites. The representative result of three independent experiments is shown (n = 2 independent samples; mean ± s.e.m.). (e) ChIP-qPCR analysis of PPARγ. Cidea 29 kb, Ppara 21 kb, Ppargc1a − 97 kb and Ucp1 9.5 kb are background sites. The representative result of three independent experiments is shown (n = 2 independent samples; mean ± s.e.m.). Source data for d,e are provided in Supplementary Table 2. (f) FAIRE-qPCR analysis. Cidea 29 kb, Ppara 21 kb, Ppargc1a − 97 kb and Ucp1 9.5 kb are background sites (mean ± s.e.m.; n = 3 independent samples; ∗∗P < 0.01). (g) RT-qPCR analysis of the indicated genes (mean ± s.e.m.; n = 3 independent samples; ∗P < 0.05, ∗∗P < 0.01). Unprocessed original scans of blots are shown in Supplementary Fig. 8.
