Figure 5.

Galangin induces autophagy and activates osteogenesis‐associated factors. A, The cells were grown on a lab‐tag chamber slide and treated with galangin (5 µM), autophagy inhibitor 3‐MA (500 µM), and autophagy inducer rapamycin (50 nM) for 24 h. The autophagosomes and autolysosomes were stained with a CYTO‐ID® Autophagy Detection Kit and analyzed by confocal microscopy. B, The expression levels of collagen I, osteocalcin, BMP‐2, ALP, Osterix, and RUNX2 were assessed using Western blot analysis. C, The gene expressions of collagen I, BMP‐2, SQSTM1/p62, and LC3A were analyzed by real‐time PCR. The data are expressed as mean ± SD (n = 3) and was analyzed by one‐way ANOVA using Dunnett's multiple‐comparison test. (*P < 0.05, **P < 0.01, and ***P < 0.001 for the difference between the control group and each treatment group)