Kartogenin Improves Osteogenesis of Bone Marrow Mesenchymal Stem Cells via Autophagy

Figure 4

KGN elevates autophagic activities in BMMSCs and prompts osteogenesis through Smad1/5/9 signal pathway. Western blotting (a) and semiquantification (b) of Smad and phosphorylation level of Smad1/5/9. 3-MA-mediated autophagy suppression inhibited KGN-induced Smad1/5/9 phosphorylation increase. , one-way ANOVA test. Western blotting (c) and semiquantification (d) of the phosphorylation level of Smad1/5/9 in the control and LDN-treated BMMSCs. LDN effectively inhibited phosphorylation level of Smad 1/5/9. , independent Student’s test. ARS detection (e) and semiquantification (f) of mineralized nodules. LDN inhibited KGN-induced osteogenesis. Scale  μm. and , one-way ANOVA test. (g) Western blotting, (h, i) semiquantification of (g), and (j, k) qRT-PCR assay of ALP and Runx2. LDN inhibited KGN-induced expression of osteogenic genes. 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). LDN inhibited KGN-induced expression of osteogenic genes. Scale  μm. , one-way ANOVA test.

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