FIG. 3.

Nucleocytoplasmic translocation of Foxo3a by the JNK pathway is independent of Akt in C2C12 cells. hFoxo3a localization was monitored by fluorescence microscopy on C2C12 cells cotransfected with vectors expressing different Akt dominant-negative (DN) HA-tagged mutants (KA, ST, and KM) and a human Foxo3a-GFP fusion expression vector (upper panel). Forty-eight hours after transfection, the cells were serum deprived for 12 h and then treated with SP600125 (20 μM) and/or anisomycin (10 μg/ml) for 1 h. Subcellular localization of Foxo3a-GFP (150 to 500 counted cells) in cells expressing Akt DN mutants is schematically represented on the graph (lower panel).