Fig 3. Effects of 25(OH)D₂ and 25(OH)D₃ on hBMSC gene expression.

Histograms depict comparative effects of 25(OH)D₂ and 25(OH)D₃ on the expression of hBMSC signature genes, opg (A), rankl (B), col1a (C), opn (D), ocn (E), vdr (F), cyp24a1 (G) and cyp27b1 (H), during their differentiation. Human BMSCs were cultured for 14 days in the presence or absence of 1000 nM of either 25(OH)D₂ or 25(OH)D₃ in differentiation media. Quantified values (RQ) have been normalised to gapdh expression and are mean ± SEM from triplicate experiments. # P<0.05, ## P<0.01 and ### P<0.001 for comparisons between 25(OH)D₂ or 25(OH)D₃ treatments and the control (vehicle); * P<0.01 for comparisons between 25(OH)D₂ and 25(OH)D₃. ND = not detectable. Western blots showing relative amounts of VDR, CYP27B1, OPN, OCN and CYP24A1 proteins in hBMSC cell lysates (20 μg protein) after 21 days of treatment with 1000 nM of either 25(OH)D₂ (D₂) or 25(OH)D₃ (D₃), or 10 nM of either 1,25(OH)₂D₂ (1D₂) or 1,25(OH)₂D₃ (1D₃) (I).