Figure 3.

Norepinephrine stimulation induces brown adipocyte-like characteristics in porcine-differentiated adipocytes. (A) Expression of brown adipogenic lineage commitment genes (nrf-1, tfam, zic1, and pgc-1α) in confluent porcine BM-MSC treated with different concentration (0–10 µM) of norepinephrine for 6 h (n = 4). The bars indicate the means ± SE. (B) Expression of common adipogenic genes (fabp4 and lpl) in differentiated adipocytes treated with 1 µM norepinephrine for 6 h (n = 4). The bars indicate the means ± SE. (C) Expression of brown fat-specific genes (prdm16, dio2, and cidea) in differentiated adipocytes treated with 1 µM norepinephrine for 6 h (n = 4). The bars indicate the means ± SE. (D) Expression of UCP proteins (UCP2 and UCP3) in differentiated adipocytes treated with 1 µM norepinephrine for 24 h (n = 3). The results shown are representative of an individual experiment. (E) Expression of UCP gene (ucp2 and ucp3) mRNA in differentiated adipocytes treated with 1 µM norepinephrine for 6 h (n = 4). The bars indicate the means ± SE. (F) Measurement of mitochondrial DNA copy number in differentiated adipocytes treated with 1 µM norepinephrine for 24 h (n = 4). The bars indicate the means ± SE. (G) Expression of mitochondria-specific genes (cox7a1, cox8b, and cyc) in differentiated adipocytes treated with 1 µM norepinephrine for 6 h (n = 4). (H) Microscographs of Oil-Red O staining in differentiated adipocytes treated with 1 µM norepinephrine (NE) for 24 h (n = 4). The results shown are representative of an individual experiment. Magnification was 60×. Bars indicate a length of 100 µm. Black arrow in control group indicates unilocular lipid droplets. Black arrow in NE-treated group indicates multilocular lipid droplets. (I) Measurement of extracellular glycerol levels in differentiated adipocytes treated with 1 µM norepinephrine for 24 h (n = 4). The bars indicate the means ± SE. *P < 0.05 vs. control. BM-MSC = bone marrow-derived mesenchymal stem cell.