Fig. 4. c-Jun phosphorylation by osteoclastogenic signals is JNK1 dependent. (A) The phosphorylation of c-Jun in BMMs was monitored by western blotting using a specific antibody directed against the phosphorylated form of c-Jun (phospho-Ser 63), 1 hour after the addition of M-CSF (20 ng/ml), RANKL (50 ng/ml), TNF- (50ng/ml) or UV irradiation (40 J/m²). (B) The expression of c-Jun was analyzed in M-CSF-(20ng/ml) and RANKL- (50 ng/ml) stimulated BMMs isolated from wild-type (wt) or Jnk1^-/- mice. (C) The activation of p38 and ERK in response to M-CSF and RANKL was analyzed by western blotting using specific anti-phosphoantibodies. (D) The activation of NF-B was analyzed by EMSA using nuclear extracts isolated from BMMs treated for 30 minutes with RANKL (+) compared with untreated cells (-).

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