Fig. 6

Sclerostin (SOST) interference ameliorates the impaired osteogenesis of BMSCs induced by AGEs via Wnt signaling. A RT-qPCR analysis of the mRNA expression of SOST in BMSCs transfected with small interfering RNAs targeting sclerostin (siSOST) or negative control (siNC) (n = 3). B RT-qPCR analysis of the expression of Runx2, Bglap and SOST in BMSCs transfected with siSOST or siNC under exposure to AGEs or not after osteogenic induction for 3 days (n = 3). C Representative images of ALP and ARS staining and quantification of ALP activity and mineralization nodules in BMSCs transfected with siSOST or siNC under exposure to AGEs or not after 7–21 days of osteogenic induction (n = 3). Scale bars = 400 μm. D Immunofluorescence assay showed the protein level and location of SOST and Runx2 in BMSCs transfected with siSOST or siNC under exposure to AGEs or not. Scale bars = 50 μm. E Western blot analysis of the expression of Runx2, Bglap and SOST in BMSCs transfected with siSOST or siNC under exposure to AGEs or not after osteogenic induction for 5 days (n = 3). F TOPflash/Renilla activity in MC3T3 cells treated with siNC, siSOST, shNC and shFTO under exposure to AGEs or not (n = 3). G Protein levels of phos-GSK3β, total GSK3β, β-catenin and SOST in BMSCs infected with shNC or shFTO under exposure to AGEs or not (n = 3). H Protein levels of Wnt/β-catenin pathway in BMSCs transfected with siNC or siSOST under exposure to AGEs or not (n = 3). Data are expressed as the mean ± SEM. ns, not significant. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001