Figure 7. Knockdown of Wnt3a or Wnt6 abolished the GYY4137 effect on Dex-treated osteoblast proliferation and differentiation. (A) All cells expressed GFP, showing that the cells were infected by lentivirus. (B–C) Verified Wnt3a or Wnt6 knockdown effect by lentivirus-mediated transduction of primary culture osteoclasts precursors. n=3, **p<0.01. (D) The proliferation of rat primary osteoblasts cells was measured by CCK8 assay after the cells were infected by lentivirus. (E) Representative photomicrographs (x200) of EdU staining and corresponding total cell photomicrographs. Blue: Hoechst labeling of cell nuclei; red: EdU labeling of nuclei of proliferative cells. (F) Quantitative data showing the percentages of EdU-positive cells in different treatment groups (number of red versus number of blue nuclei). n=3, **p<0.01. (G) Western blot analysis of Runx2 and Osterix expression in rat primary osteoblasts of each group. n=3, **p<0.01. (H) ALP staining was measured in primary osteoblast per group.