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a b c d e f Wankhade et al., Supplementary Figure 1

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1 a b c d e f Wankhade et al., Supplementary Figure 1
aP2 (Fabp4) protein expression in mature adipocytes TbRIAdKO Gapdh TbRI adipocytes SVC TbRIAdWT EWAT AWAT 8 W 16 W NC HFD EWAT Weeks on Normal Diet Body Weight (Gm) Total body weight Body weight gain Body weight and gain Minutes after glucose injection Glucose (mg/dl) Glucose tolerance test Daily food intake Daily energy intake Supplementary Figure 1 (a) aP2 (Fabp4) protein expression in mature adipocytes from EWAT and AWAT of 6 week old wild-type male mice fed normal chow (NC) or high-fat diet (HFD) for 8 weeks and 16 weeks. (b) TbRI levels in adipocytes and the SVC fraction from EWAT depots of TbR1AdKO mice and TbR1AdWT mice. The residual TbRI expression in the SVC fraction is due to presence of non-aP2-cre expressing cells that harbor intact TbRI. GAPDH levels are shown as protein loading control. (c, d) Body weight (c) and glucose tolerance (d) in TbR1AdKO (closed circle) mice and TbR1AdWT (open circle) mice fed normal chow diet over a 24 week period. (e, f) HFD food consumption in TbR1AdKO mice (closed circles) and TbR1AdWT (open circles) over (e) 2 week period (week 14 – 15) and (f) over 20 weeks of HFD feeding.

2 a b Wankhade et al., Supplementary Figure 2 EWAT UCP-1 (Abcam)
UCP-1 (Sigma) TbRIAdWT TbRIAdKO UCP-1 (Abcam) UCP-1 (Sigma) BAT BAT Supplementary Figure 2 (a, b) UCP1 antibody comparison by immunohistochemistry using Sigma and Abcam antibodies on (a) EWAT from TβRIAdWT and TβRIAdKO mice and (b) BAT from TβRIAdWT . Same ratio of primary (1:500) and secondary antibody (1:1000) were used for both antibodies.

3 a b * c Wankhade et al., Supplementary Figure 3 TbRIAdWT TbRIAdKO BAT
H&E Staining Adipocyte size in different depots TbRIAdWT TbRIAdKO BAT AWAT MWAT IWAT RWAT TbRIAdKO TbRIAdWT * c WT EWAT iBSCs stimulated with CL 316,243 Prdm16 Dio2 Ucp1 Cox8b Supplementary Figure 3 H&E staining (a) and adipocyte cell size (b) quantified in sections representing brown adipose tissue (BAT), anterior subcutaneous WAT (AWAT), mesenteric WAT (MWAT), inguinal WAT (IWAT), and retroperitoneal WAT (RWAT) in TbRIAdKO and TbRIAdWT mice on 24 weeks of HFD feeding. (c) Relative gene expression of beige/BAT specific markers in TbRIAdKO iBSCs after stimulation with CL Error Bars are expressed as +/- Standard Errors, (*p < 0.05; **p < 0.005; ***p < 0.001)

4 Thermal probe situated in graft
Wankhade et al., Supplementary Figure 4 d Thermal probe situated in graft Relative expression in recovered graft tissue compared with EWAT and BAT a TbRIAdWT TbRIAdKO b SVCs c TbRIAdWT TbRIAdKO iBSCs Supplementary Figure 4 (a) Location of the thermal probe at the site of the developed graft. The IPTT-300 transponder (BioMedic Data Systems, Seaford, DE) was sutured at the site of transplant and the local temperature was detected with a scanner (DAS-7007; BioMedic Data Systems). (b-c) H & E staining (top panels) and UCP1 staining (bottom panels) in graft sections obtained from TbRIAdKO and TbRIAdWT SVCs (b) and iBSCs (c) after cold exposure (see details in methods section). (d) Individual gene expression levels from two transplanted mice for classic beige/BAT specific markers in EWAT, BAT and transplanted grafts from iBSCs (referred to as ADSCs here) harvested from TbRIAdKO and TbRIAdWT mice. Error bars are expressed as +/- Standard Errors, (*p < 0.05; **p < 0.005; ***p < 0.001).


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