Figure 2
Effects of orobol on 3T3-L1 preadipocytes at different stages of MDI-induced adipogenesis. (A) A time schedule for orobol treatment during cellular differentiation. (B,C) Confluent 3T3-L1 preadipocytes were differentiated into mature adipocytes in the presence of 20 μM orobol for the indicated time periods. (B) After differentiation, the images of 3T3-L1 adipocytes stained with Oil Red O solution were captured. (C) Stained lipids were quantified via spectrophotometry as described in the Materials and Methods. The sharps (##) indicate a significant difference between the control group and the group treated with MDI alone (p < 0.01). Means with different letters (a–f) within a graph are significantly different between groups treated with MDI alone and those treated with MDI plus orobol treated at each of the different stages. (D) Orobol delays MDI-stimulated cell cycle progression at the G1 phase. When 3T3‐L1L preadipocytes reached confluence, orobol (20 μM) or daidzein in DMEM supplemented with 10% FBS containing MDI was added, and the extent of cell proliferation examined. After the indicated time periods (16 h), cells were dissociated with trypsin/EDTA and FACS analysis was conducted. Data are representative of three independent experiments that yielded similar results. Data are represented as mean ± SEM from at least three independent experiments. Means with different letters (a–c) within a graph are significantly different between groups.
