Fig. 6.

HOXD8 initiates RUNX2 expression to drive osteogenic differentiation. a HOXD8 together with RNA polymerase II (Pol II) and H3K4me3 were highly enriched on RUNX2 promoter by ChIP assays. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. b BM-MSCs were infected with indicated lentivirus and RUNX2 expression was assessed by qPCR. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. c Normal BM-MSCs were infected with indicated lentivirus and cultured in MSC media for 3 days. Indicated BM-MSCs were cultured as in Fig. 2e. Expression levels of self-renewal related genes were assessed by real-time qPCR. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. d Normal BM-MSCs were infected with indicated lentivirus and cultured in MSC media for 3 days. Indicated BM-MSCs were cultured as in Fig. 2f. Expression levels of osteogenic differentiation genes were assessed by real-time qPCR. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. e Normal BM-MSCs were infected with indicated lentivirus and cultured in MSC media for 3 days. Osteogenic differentiation was induced as in Fig. 2f. ALP staining was performed on day 6 of osteogenic differentiation. f Normal BM-MSCs were infected with indicated lentivirus and cultured in MSC media for 3 days. Osteogenic differentiation was induced as in Fig. 2f. Von Kossa staining was performed to indicate mineral deposition on day 12. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. Scale bar, 10 μm. Color intensity of mineral deposition was quantified by Image J. Intensity changes were counted as means ± S.D. **P < 0.01. g Indicated BM-MSCs were infected with lentivirus and cultured as in Fig. 2f. ALP staining was performed on day 6 of osteogenic differentiation. h Indicated BM-MSCs were infected with lentivirus and cultured as in Fig. 2f. Von Kossa staining was performed to indicate mineral deposition on day 12. Data are from three independent experiments using BM-MSCs derived from 3 healthy donors. Scale bar, 10 μm. Color intensity of mineral deposition was quantified by Image J. Intensity changes were counted as means ± S.D. **P < 0.01