Figure 8.
PKCζ is a downstream target of Phlpp1 that controls osteoclastogenesis and bone resorption. Bone marrow macrophages were collected from female Phlpp1 cKOCtsk and Cre+ control littermates. Cells were cultured in the presence of 60 ng/ml of RANKL and 25 ng/ml of M-CSF. On day 3, cells were exposed to a PKCζ pseudosubstrate for 24 h and (A) phalloidin Alexa 488, DAPI, and phase-contrast images were collected. TRAP staining was performed (B) and the number of osteoclasts was determined (D). *, p < 0.05. C, cells were seeded onto bovine bone slices and cultured in the presence of 60 ng/ml of RANKL and 25 ng/ml of M-CSF for 14 days in the presence of the PKCζ pseudosubstrate or vehicle control (PBS). E and F, resorption pits were visualized by toluidine blue staining and the number of pits (E) and percent of each bone slice resorbed (F) was evaluated using ImageJ software. *, p < 0.05.