FIG. 5.

EPO promotes brown features in WAT. A: CS activity in various primary adipocytes isolated from WT control and EpoR^aP2KO mice (KO) without (□, PBS) or with (■) EPO treatment was determined (n = 4). B: Hematoxylin and eosin staining of S-WAT and V-WAT from WT and EpoR^aP2KO (KO) mice is shown (scale bar: 20 μm). C: Immunohistochemistry staining for UCP-1 in S-WAT and V-WAT from WT and EpoR^aP2KO (KO) mice is shown (scale bar: 20 μm). SVF from the inguinal fat pad was differentiated into adipocytes for 6 days (M-adipocytes) without (PBS) and with EPO treatment (5 units/mL), and expression of BAT associated genes (D) and mitochondrial biogenesis–related genes (E) was determined using quantitative PCR. F: Mitochondria and the nucleus were visualized using MitoTracker (red) and DAPI (blue), respectively, and confocal microscopy. The white arrows indicate lipid droplets. Cells were counterstained with DAPI to detect nuclei (blue). Views are at original magnification ×60. Scale bars: 10 μm. G: Total and uncoupled OCRs in M-adipocytes treated without (PBS) or with EPO (5 units/mL) were monitored. Data are representative of three independent experiments and normalized to total cellular protein. Statistics were performed using the Student t test. Bar graphs are mean ± SEM. Data are averages of three independent experiments. Ctrl, control; OM, oligomycin. *P < 0.05; **P < 0.01.