Kartogenin Improves Osteogenesis of Bone Marrow Mesenchymal Stem Cells via Autophagy

Figure 5

BMMSC osteogenic level promoted by KGN is suppressed by 3-MA, whereas rescued by SB4. Western blotting assay (a) and semiquantification (b) of Smad1/5/9 and phosphorylation level of Smad1/5/9 in the control and SB4-treated BMMSCs. SB4 promoted phosphorylation level of Smad1/5/9. , independent Student’s test. Western blotting assay (c) and semiquantification (d) on Smad1/5/9 and phosphorylation level of Smad1/5/9. 3-MA suppressed KGN-induced increased phosphorylation level of Smad1/5/9, whereas SB4 reversed this effect. and , one-way ANOVA test. ARS (e) and semiquantification (f) of mineralized nodules. 3-MA restrained KGN-induced BMMSC osteogenesis, whereas SB4 reversed this effect. Scale  μm. and , one-way ANOVA test. (g) Western blotting, (h, i) semiquantification of (g), and (j, k) qRT-PCR of ALP and Runx2. 3-MA suppressed KGN-induced expression of osteogenic genes whereas SB4 reversed this effect. , , and , one-way ANOVA test. (l, m) Detection of ALP (green), Runx2 (green), and DAPI (blue) by immunofluorescent staining in BMMSCs. (n, o) Semiquantification of (l) and (m). Scale  μm. and , one-way ANOVA test.

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