Figure 3

BRWE promotes browning of adipocytes in 3T3-L1 adipocytes. Two days after achieving confluence, 3T3-L1 pre-adipocyte cells were stimulated by DM (Be) containing 0.5 mM IBMX, 167 μM insulin, T3 20 μM, and 0.5 μM troglitazone with/without various concentrations of BRWE. After 6 days, intracellular lipid droplets were measured with (A,B) Oil-Red O assay and the absorbance was measured at 500 nm. (C) The mRNA and (D,E) protein levels of thermogenesis-related factors in 3T3-L1 with BRWE were measured. (F) Expression of intracellular UCP1 and PGC1α in 3T3-L1 were evaluated by immunofluorescence staining. The changes were observed at 200× (scale bar = 100 μM). Florescent intensities were quantified using the ImageJ software and presented as a graph in arbitrary units. (G) Mitotracker staining was performed and observed at 200× (scale bar = 150 μM). (H) Protein levels of NRF1, CIDEA, and CPT1B were measured. All data are expressed as the mean ± SEM. ^#p < 0.05, significantly different from DM (Be)-untreated pre-adipocytes; * p < 0.05 and ** p < 0.01, significantly different from DM (Be)-stimulated adipocytes cells. Each experiment was repeated at least 3 times, and similar results were obtained. BRWE, black raspberry water extract.