FIGURE 5.

AKT phosphorylation and activity is increased in Cbl^YF/YF OCLs. A, OCLs were serum-starved for 30 min and treated with RANKL (50 μg/ml) for the indicated times. The blot was probed with anti-phospho-Cbl Tyr⁷³⁷ (pY737Cbl) antibodies to detect the phosphorylation of Cbl Tyr⁷³⁷ (top). The blot was stripped and reprobed with anti-Cbl antibodies to detect total Cbl levels in WT and Cbl^YF/YF OCLs. The amounts of total proteins in individual bands were quantified as described above. B, OCLs were treated with RANKL as described above. Blots were probed with anti-phospho-AKT Thr³⁰⁸ (pAkt) (top) and anti-AKT antibodies (bottom). Quantification of blots was performed as above. C, phosphorylation of GSK is enhanced in Cbl^YF/YF OCLs. OCLs were treated with RANKL as described above. Western blot was performed, and the blots were probed with anti-phospho-GSK and anti-GSK antibodies. Quantification of blots was performed as above.