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. 2021 Jul 25;17(12):3173–3187. doi: 10.7150/ijbs.62556

Figure 4.

Figure 4

The regulation of adipose tissue remodeling and thermogenesis by BBR is dependent on SIRT1. After 8 days of differentiation, 3T3-L1 and HIB1b cells were treated with vehicle or BBR (5 μM) for 24 h. (A) The fully differentiated cells were fixed and subjected to Mito-tracker Red staining, Scale bar=100μm. (B) Representative transmission electron microscopy images from fully differentiated 3T3-L1 cells treated with vehicle and BBR (5 μM), Scale bar=1μm (×12000). L: lipid droplets, M: mitochondria. The proteins of SIRT1, PPARγ, and UCP1 were expressed in fully differentiated (C) 3T3-L1 and (D) HIB1b cells treated with vehicle or BBR (0.5 μM and 5 μM, respectively) for 24 h. 3T3-L1 and HIB1b cells with stable knockdown SIRT1 were established by lentivirus transfection. (E) 3T3-L1 and (F) HIB1b cells transfected with SIRT1 plasmid for 36 h or treated with BBR for 24 h. SIRT1 knockdown 3T3-L1 cells were treated with or without BBR for 24 h. The expressions of SIRT1, PPARγ, and UCP1 were detected by Western blot. (G) Relative mRNA levels of thermogenesis genes and fatty acid oxidation genes in NC or SIRT1 knockdown 3T3-L1 cells after differentiation treated with vehicle or BBR 12 h. *P<0.05 and **P<0.01 compared with NC-Veh, ##P<0.01 compared with NC-BBR. (H) After full differentiation, 3T3-L1 and shSIRT1 3T3-L1cells were treated with or without BBR for 24 h and stained Oil red O.