Fig. 7

Shikonin alleviates the inhibition of osteogenic function of BMSCs by activating the AMPK pathway. (A, B) ALP and ARS staining results after 7 and 14 days of osteogenic induction using shikonin. Semiquantitative analysis of ALP staining and calcium deposition is shown in the bar charts. (scale bar: 100 μm). (C, D) Effect of shikonin on osteogenesis-related genes and proteins of BMSCs exposed to the HG inflammatory environment or not. Full-length blots are presented in Supplementary Fig. S5. (E) Western blot analysis showing AMPK pathway activation following shikonin treatment. Full-length blots are presented in Supplementary Fig. S5. (F, G) Western blot bands representing the effects of an AMPK activator and inhibitor (F), and the correlation between AMPK activation and osteogenesis (G). Full-length blots are presented in Supplementary Fig. S5. (H, I) ALP staining and ARS staining images with semi-quantitative analysis. (scale bar: 100 μm). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; ns, not significant