Figure 4

Expression of early, middle, and late-stage lineage-specific osteogenic genes in bone marrow-derived mesenchymal stem cells was significantly induced by daidzein analogs. Bone marrow-derived mesenchymal stem cells (BMSCs) 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 to vehicle-treated BMSCs; ***P < 0.001 compared with vehicle-treated BMSCs; ^#P < 0.05 compared with E2-treated BMSCs; ^###P < 0.001 compared with E2-treated BMSCs; ^ΨP < 0.05 compared with daidzein-treated BMSCs; ^ΨΨΨP < 0.001 compared with daidzein-treated BMSCs.