(A) Bcl-xfl/fl osteoclasts were infected
with AxGFP, AxCre, AxBcl-xL, AxMekCA, or AxRasDN. Bcl-xL
overexpression by AxBcl-xL infection suppressed, and knockout of
Bcl-x gene by AxCre infection increased, Erk activity, as
determined by the amount of phospho-Erk in Bcl-xfl/fl
osteoclasts. In contrast, AxMekCA infection increased, and
AxRasDN infection decreased, Bcl-xL expression. Relative intensity of
the bands on each gel, measured by densitometry, is shown above each lane.
(B) Bcl-xfl/fl osteoclasts were
infected with AxGFP, AxGFP plus AxRasDN, AxBcl-xL, or AxBcl-xL plus
AxRasDN. Reduced osteoclast survival by RasDN
overexpression was completely rescued by Bcl-xL overexpression.
*P < 0.01 versus AxGFP-infected cells. (C)
Bcl-xfl/fl osteoclasts were infected with AxGFP
or AxBcl-xL, and then treated with the indicated concentrations of MEK inhibitor
PD98059. PD98059 treatment dose-dependently suppressed the survival of
osteoclasts, which was completely rescued by Bcl-xL overexpression.
*P < 0.01 versus untreated osteoclasts. (D)
Bcl-xfl/fl osteoclasts were infected with AxGFP
or AxCre together with AxMekCA. Prosurvival effect of MekCA
overexpression was partially suppressed by Bcl-x deletion.
*P < 0.01, **P < 0.05 versus
AxGFP-AxMekCA–infected cells. All results are mean
± SD of 6 cultures.