Figure 3.

Functions of Wnt5a and Wnt5b in SMSC-Exos-stimulated articular chondrocytes. (A) Normalized gene expression levels of Wnt family members in SMSCs. This experiment used three replicates. (B) Expression of YAP downstream genes in chondrocytes treated with exosomes, at a concentration of 10 × 10¹¹ exosome particles/mL, derived from SMSCs transfected by empty vector (SMSC-Exos-10) and SMSCs transfected with shWnt5a and shWnt5b (SMSC-shWnt5a&5b-Exos). This experiment was repeated three times. (C) YAP activation was detected by western blotting by observing the de-phosphorylation of YAP. The results of statistical analysis of three independent replicates are also shown. *P < 0.05 compared to control. (D) Subcellular localization of YAP was observed using confocal immunofluorescence microscopy. This experiment was repeated independently three times and representative results are shown. Scale bar: 50 μm. (E) EdU assays were performed using flow cytometry. The percentage of EdU-positive cells (labelled green) was determined. This experiment was repeated independently three times and representative results are shown. (F) Migration assays of chondrocytes. Migrated chondrocytes were stained by crystal violet. This experiment was repeated independently three times and representative results are shown. Scale bar: 50 μm. (G) Gene expression changes of Aggrecan, Col II and SOX9 after stimulation with the indicated exosomes, at a concentration of 10 × 10¹¹ exosome particles/mL. This experiment was repeated three times. *P < 0.05 compared to control. (H) Protein expression levels of Col II, Aggrecan and SOX9 were detected using western blotting. The results of statistical analysis of three independent replicates are also shown. *P < 0.05 compared to control.