Fig. 3

Risperidone confers an inhibitory effect on differentiation and autophagy and a promotive effect on apoptosis of osteoblasts in vitro. A the mRNA expression of OPG, collagen I and RANKL in MC3T3-E1 cells determined by RT-qPCR. B–C Western blot analysis of differentiation-related protein (OPG, collagen I and RANKL) expression in MC3T3-E1 cells. The band intensity was assessed. D representative images of ALP staining in MC3T3-E1 cells. E, statistical results of ALP positive rate in MC3T3-E1 cells. F representative images of alizarin red S staining in MC3T3-E1 cells. G statistical results of integrated optical density (IOD) of alizarin red S in MC3T3-E1 cells. H representative images of autophagosome in MC3T3-E1 cells assessed by immunofluorescence. I Western blot analysis of autophagy-related protein (LC3 II/I, Beclin1, and p62) expression in MC3T3-E1 cells. The band intensity was assessed. J Western blot analysis of apoptosis-related protein (cleaved PARP1, cleaved caspase-3, cleaved caspase-8 and cleaved caspase-9) expression in MC3T3-E1 cells. K apoptosis rate in MC3T3-E1 cells evaluated by flow cytometry. *p < 0.05, compared with MC3T3-E1 cells treated with normal culture medium. The cell experiments are repeated 3 times