Figure 6.

Phlpp1 deficiency increases ex vivo osteoclastogenesis. Bone marrow macrophages were collected from female Phlpp1^−/− or WT littermate 6-week-old mice. A, cells were cultured in the presence of 60 ng/ml of RANKL and 25 ng/ml of M-CSF for 3 days. Cultures were then serum starved in the absence of RANKL and M-CSF for 1 h. Following serum starvation, 25 ng/ml of M-CSF was added back to cultures for the indicated times (minutes) and Western blotting was performed. B, Western blotting was performed using bone marrow macrophages. C, cells were cultured in the presence of 30 ng/ml of RANKL and 25 ng/ml of M-CSF for 4 days and c-fms immunofluorescence was performed.