Fig. 6.

ActA decreases CatK release from osteoclasts. Mature murine osteoclasts (day 10) were treated with MCSF alone (20 ng/ml) or in combination with RANKL (50 ng/ml) and ActA (50 ng/ml) in the presence of bovine bone powder. Conditioned medium (CM) and total cell lysates (TCL) were collected at 24, 48 and 72 hours post-treatment initiation. (A) Western blot of 50 µg of total protein from conditioned medium and TCL identifying CatK and β-tubulin, which was used to normalize and account for differences in lane loading. Western blots were visualized using the LI-COR Odyssey Quantitative western blotting system. (B) Ratio of conditioned medium CatK to total CatK, first normalized to the TCL β-tubulin levels to account for differences in lane loading. Values are mean±s.d. calculated from two independent experiments. Different letter designations indicate significantly different groups within each time point (P<0.05).