Figure 5

Expression of early, middle, and late-stage lineage-specific osteogenic genes in adipose-derived stromal/stem cells was significantly induced by daidzein analogs. Adipose-derived stromal/stem cells (ASCs) were cultured in charcoal dextran-stripped osteogenic differentiation medium and concurrently treated with vehicle, 17β-estradiol (E2; 10 nM), daidzein (1 μM), or daidzein analog 2g or 2l (1 μM). Cells were collected after 3, 7, or 14 days of treatment. RNA was isolated from the cells and reverse transcribed into cDNA. Analysis of (A) early, (B) middle, or (C) late osteogenic transcription factors by quantitative polymerase chain reaction. Expression values are normalized to undifferentiated cells normalized to 1.0. Bars, ± standard deviation. *P < 0.05 compared with vehicle-treated ASCs; **P < 0.01 compared with vehicle-treated ASCs; ***P < 0.001 compared with vehicle-treated ASCs; ^###P < 0.001 compared to E2-treated ASCs; ^ΨΨΨP < 0.001 compared with daidzein-treated ASCs.