Fig. 3.

Adipocyte-autonomous role for GRAF1 in BAT formation and function. (a) differentiation of primary pre-brown adipocytes isolated from the stromal vascular fraction (SVF) of iBAT was reduced by GRAF1 depletion as assessed by UCP1 expression. n = 3–4. (b,c). Oil red O staining of differentiated SVF cells treated with indicated siRNA (b) and quantification of oil red O staining (c). (siG1 denotes siGRAF1). n = 3/treatment. (d) Assessment of GRAF1 depletion in BAT and scWAT in female WT and GRAF1^AKO mice via Western blot analysis. e. Female GRAF1^AKO mice exhibited cold intolerance when subjected to conditions described in (Fig. 2f). n = 5–6/group. (f) qRT-PCR data showed significantly decreased markers of brown adipogenesis, mitochondrial components and FGFs in iBAT of GRAF1^AKO mice. n = 6/group. Data are represented as mean ± SEM, *P < 0.05; **P < 0.01; ***<0.001;****P < 0.0001 by two-tailed student’s t-test (a, c,f) or repeated measures ANOVA with post-hoc Fisher’s LSD test (e).