## 2 Materials and Methods
### Antioxidant Activity  
In the DPPH radical-scavenging activity assay, 0.2 mL of each of the BR, BRB, and MBR solutions was mixed with 0.8 mL of a 0.0004 M DPPH solution. The reaction mixture was then incubated in the dark for 10 min. The absorbance of DPPH was measured at 517 nm, and the radical-scavenging activity was calculated as follows (2):  
\[\text{DPPH radical-scavenging activity (\%)}=(1-\text{Aexperiment/Acontrol})\times 100 \tag{2}\]  
For the ABTS radical-scavenging activity assay, 7 mM ABTS and 2.45 mM potassium persulfate solutions were mixed and incubated in the dark for 16 h to allow free radical generation. The mixture was then diluted to obtain an absorbance of 0.70 \(\pm\) 0.02 at 734 nm. Afterward, 1 mL of the ABTS solution was mixed with 10 \(\upmu\)L of each of the BR, BRB, and MBR solutions. The resulting mixture was incubated in the dark for 6 min. The absorbance of ABTS at 734 nm was measured, and the radical-scavenging activity was calculated as follows (3):  
\[\text{ABTS radical-scavenging activity (\%)}=(1-\text{Aexperiment/Acontrol})\times 100 \tag{3}\]  
For the ferric-reducing antioxidant power (FRAP) assay, an FRAP reagent was prepared by mixing 300 mM sodium acetate buffer (pH 3.6), 10 mM 2,4,6-tripyridyl-s-triazine (TPTZ), and 20 mM FeCl\({}_{3}\)-6H\({}_{2}\)O (ferric chloride) at a ratio of 10:1:1. Then, 50 \(\upmu\)L of each of the BR, BRB, and MBR extracts, 1.5 mL of the FRAP reagent, and 150 \(\upmu\)L of DW were mixed and incubated at 37 \({}^{\circ}\)C for 4 min. The absorbance of the resulting mixture was measured at 593 nm. Evaluation of the Effects of 80% EtOH Extracts of BR, BRB, and MBR on Cell Viability and Lipid Accumulation in Obesity and Obesogenic Models  
3T3-L1 cells were obtained from the American Type Culture Collection (ATCC, CL-173; Manassas, VA, USA) and were cultured in 24-well and 96-well plates in DMEM containing 10% BS, 3.7 g/L sodium bicarbonate, and 1% P/S in a 37 \({}^{\circ}\)C/5% CO\({}_{2}\) incubator until confluence. Two days later, 100% confluence was reached (day 0); the preadipocytes were cultured in 10% FBS medium with adipocyte differentiation-inducing substances (1.0 \(\upmu\)g/mL insulin, 1.0 \(\upmu\)M DEX, and 0.5 mM IBMX; MDI) to induce cell differentiation. After two days, the medium was changed to a 10% FBS medium containing 1.0 \(\upmu\)g/mL of insulin to promote further cell differentiation. The medium was changed every two days, and 3T3-L1 adipocytes were obtained after 8 days of treatment. The viability of adipocytes treated with 80% EtOH extracts of BR, BRB, and MBR was evaluated using the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazol-5-carboxanilide (XTT) assay kit (WelGene, Seoul, Republic of Korea). On day 8 of differentiation, a mixture of 1 mL XTT and 20 \(\upmu\)L N-methyl dibenzopyrazine methyl sulfate (PMS) was prepared and added to each well of the 96-well plate. The plate was then incubated for 4 h at 37 \({}^{\circ}\)C in a 5% CO\({}_{2}\) incubator. The absorbance was measured at 450 and 690 nm. Thereafter, lipid accumulation from 400 \(\upmu\)g/mL of 80% EtOH extracts of BR, BRB, and MBR on 3T3-L1 cells was measured. The culture solution was aspirated from the 24-well plates and washed twice with PBS. Then, 500 \(\upmu\)L of a 10% formalin solution was added to each well and incubated at room temperature for 1 h to fix the differentiated adipocytes. The cells were washed with 500 \(\upmu\)L of a 60% isopropanol solution and dried completely at room temperature. The completely dried cells were stained with 200 \(\upmu\)L of an ORO solution added to each well and incubated for 30 min. After washing with DW, the cells were completely dried at room temperature for 1 day. The ORO combined lipids were eluted with 100% isopropanol, and the absorbance was measured at 490 nm. Based on the above methods, the anti-obesity efficacy of the 80% EtOH extracts of BR, BRB, and MBR was evaluated.