Figure 7. Over-expressed Klotho binds to Wnt and blocks Wnt-mediated gene transcription in vitro. Chondrocytes were incubated with or without CTS(1 HZ) for 48 hours. Overexpression of klotho attenuated the activity of the wnt signaling pathway (A) Co-IP method was used to detect the binding of Klotho and Wnt1, Wnt4 and Wnt7a in different groups of cells supernatant. (B) Co-IP method was used to detect the binding of Klotho and Wnt1, Wnt4, Wnt7a, the binding of Dvl to Frizzled in different groups of cells, quantitative data (C, D) is presented. *P,0.05 versus controls (n=3), The cell lysates were immunoprecipitated by β-catenin antibody. qPCR analyses the enrichment of MMP-2, MMP-9-specific DNA in chondrocytes. (E, F) Overexpression of klotho blocks the binding of β-catenin on MMP-2 and MMP-9 promoter activity. Results were obtained via the expression of three individual experiments performed in triplicate for each condition. *P < 0.05, **P < 0.01.