FIGURE 5.

M-CSF induces Cbl phosphorylation and NFATc1 degradation. A, preosteoclasts were starved and pretreated with vehicle or SU6656 (2 μm) for 30 min before stimulation with M-CSF as indicated. Lysates were immunoprecipitated (IP) with anti-Cbl-b or anti-c-Cbl antibodies. Immunoprecipitated samples were probed with anti-phosphotyrosine 4G10, anti-Cbl-b, or anti-c-Cbl antibodies. B, BMMs were cultured for 2 days with M-CSF and RANKL. Cultured cells were washed and further incubated with M-CSF alone or M-CSF and RANKL. MG132 (25 μm) was added 3 h before cell lysis. Lysates were probed with anti-NFATc1 and control anti-actin antibodies.