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. 2018 Jun 7;96(9):3635–3644. doi: 10.1093/jas/sky230

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Figure 2. — BMP7 promotes brown preadipocyte commitment and terminal differentiation of porcine BM-MSC. (A) Expression of bone morphogenetic protein receptor genes (bmpr1a, bmpr1b, and bmpr2) in confluent porcine BM-MSC (n = 4). The bars indicate the means ± SE. (B) Expression of brown adipogenic lineage commitment genes (nrf-1, tfam, zic1, and pgc-1α) in porcine BM-MSC treated with BMP-7 for 3 d (n = 4). The bars indicate the means ± SE. (C) Oil-Red O staining of porcine BM-MSC after stimulation of differentiation by a brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). The results shown are representative of an individual experiment. (D) Quantification of intracellular triglyceride levels in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). (E) Expression of common adipogenic genes (fabp4 and lpl) in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). The bars indicate the means ± SE. (F) Microscographs of Oil-Red O staining of porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). Magnification was 60×. Bars indicate a length of 100 µm. Black arrow in control group indicates unilocular lipid droplets. Black arrow in BMP7-treated group indicates multilocular lipid droplets. The results shown are representative of an individual experiment. (G) Expression of brown fat-specific genes (prdm16, dio2, and cidea) in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). The bars indicate the means ± SE. (H) Expression of UCP proteins (UCP2 and UCP3) in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 3). The results shown are representative of an individual experiment. (I) Expression of UCP gene (ucp2 and ucp3) mRNA in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). The bars indicate the means ± SE. (J) Measurement of mitochondrial DNA copy number in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). The bars indicate the means ± SE. (K) Expression of mitochondria-specific genes (cox7a1, cox8b, and cyc) in porcine BM-MSC treated with brown adipogenic induction cocktail for 14 d (BMP7 + MDI) (n = 4). (L) The extracellular glycerol levels in brown adipogenic induction cocktail (BMP7)-treated porcine BM-MSC in response to 1 µM norepinephrine (NE) treatment for 6 h (n = 4). The bars indicate the means ± SE. (F) *P < 0.05 vs. control. BM-MSC = bone marrow-derived mesenchymal stem cell.