Fig. 5.

SDF-1 mediates increased migration but not differentiation of osteoclast precursors. Transmigration assay of osteoclast precursors in response to SDF-1. A. Osteoclast precursors were placed in the upper chamber of the transwell with or without AMD3100. SDF-1 was added to the lower chamber. Percentage of cells, which migrated to the lower chamber was quantified and data is expressed relative to WT cells without treatment in the absence of SDF-1 in the lower chamber. B. Osteoclast precursors were differentiated in the presence of MCSF (20 ng/ml) and RANKL (50 ng/ml) in the presence or absence of SDF-1 (100 ng/ml) added at indicated times. After 5 days of differentiation, cells were TRAP stained and TRAP positive cells with >3 nuclei were considered as osteoclasts. C. Osteoclast differentiation was performed in the presence or absence ofAMD3100 with SDF-1 treatment (100 ng/ml). Images of TRAP positive cells at 10× magnification are shown. D. Bar graph shows the number of multinucleated (>3 nuclei/cell) TRAP positive cells. In panel A, a: compared to untreated WT cells, b: compared to WT cells exposed to SDF-1 in lower chamber, c: compared to YF cells exposed to SDF-1 in lower chamber. In B, D * compared to WT cells without SDF-1 or AMD3100 treatment. A representative of 3 independent experiments is shown. * P value of <0.05 from ANOVA, Tukey's post hoc test was considered statistically significant.