Fig. 4

Wnt5a promotes osteoclast formation through enhanced dendrocyte-expressed seven transmembrane protein (DCSTAMP) and matrix metalloproteinase 9 (MMP9) expression. Osteoclasts were formed in vitro from bone marrow-derived monocytes in L cell conditioned medium (Control) or L Wnt5a conditioned medium (Wnt5a). Osteoclasts were visualized after 5 days of RANKL stimulation by tartrate-resistant acid phosphatase (TRAP) (dark color) and 4',6-diamidino-2-phenylindole (a). Osteoclasts were enumerated by identifying TRAP+ cells with three or more nuclei and expressed as frequency per 1000 cells or by percent of nuclei in osteoclasts (b). By both measures, Wnt5a increased the osteoclast formation (**p ≤ 0.01, ***p ≤ 0.001). c mRNA isolated from fusing osteoclast cultures at day 2 and day 5 were analyzed for DCSTAMP and MMP9 expression. The levels were increased by Wnt5a treatment. *P ≤ 0.05 for comparison of control and Wnt5a-treated cultures