Fig. 4

c-Cbl interacted with ubiquitinated TRAF6 after stimulation with RANKL and IFN-γ. a – Osteoclasts differentiated from bone marrow precursors were stimulated with RANKL for the indicated times and lysed. Cell extracts were immunoprecipitated with anti-cbl Ab and then probed with anti-TRAF6. The presence of endogenous TRAF6 or c-Cbl was analyzed via western blotting using anti-TRAF6 and anti-cbl. b – BMMs were extracted, as described in the Methods section, and treated with M-CSF (30 ng/ml). After three days, cells were stimulated with IFN-γ as indicated. Cell extracts were immunoprecipitated with anti-cbl Ab and then probed with anti-TRAF6. The presence of endogenous TRAF6 or c-Cbl was analyzed via western blotting using anti-TRAF6 and anti-cbl, as indicated