FoxO1 and FoxO3a knockdown partially restores osteogenic transcription factor OSX and chondrogenic transcription factor SOX9 expression. A, Relative FoxO1 mRNA expression in mesenchymal stem cells (MSCs) following nontargeting, FoxO1, or FoxO3a siRNA knockdown. B, Relative osterix (OSX) mRNA expression in differentiating MSCs treated with 50 mM EtOH or media alone for 24 hours. C, Relative SRY-related high-mobility group-box 9 (SOX9) mRNA expression in differentiating MSCs treated with 50 mM EtOH or media alone for 48 hours. D, Relative FoxO3a mRNA expression in MSCs following nontargeting, FoxO1, or FoxO3a siRNA knockdown. E, Relative OSX mRNA expression in differentiating MSCs treated with 50 mM EtOH or media alone for 24 hours. F, Relative SOX9 mRNA expression in differentiating MSCs treated with 50 mM EtOH or media alone for 48 hours. Media control and differentiation control (first and second columns in panels B, C, E, and F) are shown to confirm differentiation marker expression and are not for statistical purposes. Different letters demonstrate significance, n = 4, p < 0.05,1-tailed Student’s t-test.