Fig. 3

NAD⁺ is indispensable for BMSC osteogenic commitment. A Intracellular level of NAD⁺, NADH, and the ratio of NAD⁺/NADH in hBMSCs at day 7. B Relative expression of NAMPT in hBMSCs at day 7. C Representative image of western blot analysis with the hBMSCs at day 7. Quantification of the western blot from three independent experiments was shown upon the bands. N = 3. D Intracellular level of NAD⁺ of hBMSCs with vehicle (DMSO), FK866, or P7C3 treatment for 7 days. E Representative image of ALP staining (top) and Alizarin Red staining (bottom) for hBMSCs with DMSO, FK866, or P7C3 treatment. The staining of hBMSCs at D0 was presented as control. Right panels were quantification of the ALP or alizarin red staining from four independent experiments. N = 4. F Relative expression of osteogenic-related genes RUNX2, ALP, SP7, COL1α1, BGLAP and SPP1 in hBMSCs with P7C3 or FK9866 treatment at day 3 and day 7. DMSO group at day 3 was used as control. N = 3. Statistical significance was calculated with one-way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001. Data are represented as mean ± SD