FIGURE 6.

Cbl proteins mediate NFATc1 degradation. A, BMMs from WT and Cbl-b-deficient mice were cultured with M-CSF and RANKL for the indicated times. Lysates were immunoblotted with NFATc1 and actin antibodies. KO, knock-out. B and C, BMMs from WT and Cbl-b-deficient mice were transduced with pSuper-Retro (control) or a c-Cbl siRNA-expressing (c-Cbl-si) retrovirus, respectively. Cells were cultured with M-CSF and RANKL for the indicated times. B, lysates were probed with NFATc1, Cbl-b, c-Cbl, and actin antibodies (upper panel). The relative amounts of NFATc1 are shown in the lower panel. C, real-time PCR was performed for detection of NFATc1. B and C, ns, not significant, *, p < 0.01 versus positive control. Data are expressed as mean ± S.D. of triplicate samples. Results presented are representative of three independent sets of similar experiments.