Fig. 6

Effect of 3-MA on the differentiation and mineralization of osteoblasts by kaempferol. a The MC3T3-E1 cells were pretreated with an autophagy inhibitor, 3-MA (1 mM), for 1 h and then treated with 10 μM of kaempferol. ALP activity showed that 3-MA blocked the differentiation of the MC3T3-E1 cells induced by the kaempferol (10 μM) treatment. The data are expressed as the mean ± SD (n=6). The statistical significance of each group was analyzed by a one-way ANOVA and Tukey’s HSD test (*p < 0.001 [3 days] and #p < 0.001 [7 days] for the difference between the control compared to the treatment group). b 3-MA also inhibited osteoblast mineralization induced by the addition of kaempferol, as shown by alizarin red staining. The data are expressed as the mean ± SD (n=6). The statistical significance of each group was analyzed by a one-way ANOVA and Tukey’s HSD test (*p < 0.001 [7 days] and #p < 0.001 [14 days] for the difference between the control group compared to the treatment groups)