Localization of Mdm2 characterized by confocal microscopy.
(a) Endogenous Mdm2 in serum-starved MCF-7
cells transfected with constitutively active Akt (CAAkt) or incubated
in the absence or presence of LY294002 and then treated with IGF-1 (5
min). (b) HEK 293 cells transiently transfected with
human Flag-Mdm2 and Flag-Mdm2, then treated with wortmannin (1 h),
Flag-Mdm2 and KD-Akt, Flag-Mdm2 and Δp85 PI3-kinase, and Flag-Mdm2
and KD-Akt, and then treated with IGF-1. (c) Primary,
serum-starved human keratinocytes were treated with growth factor-free
medium (control) or medium containing IGF-1. Keratinocytes were also
transfected with Flag-Mdm2, Flag-Mdm2, and KD-Akt, or treated with
LY294002 (1 h). (d) HEK 293 cells transfected with
Flag-Mdm2, Flag-S166A, Flag-186A, and Flag-2xSA. Additionally, cells
were cotransfected with Flag-2xSA and CA-Akt or were transfected with
Flag-2xSA and then treated with IGF-1.