FIGURE 5.

DGKδ and PKCα modulate Akt phosphorylation and USP8 levels. A, DGKδ and/or PKCα were depleted in HeLa cells using RNAi and then the levels of phosphorylated MARCKS (pMARCKS), MARCKS, PKCα, DGKδ, and actin were determined by Western blotting. Lanes were removed from the blot for clarity. The densities of the pMARCKS bands (arbitrary units) are indicated above the blot. B, HeLa cells were transfected with scrambled or DGKδ siRNA duplexes, starved for 16 h, exposed to 250 nm Gö6976 and then to EGF (10 ng/ml) for 10 min. The levels of the indicated proteins in cell lysates were detected by Western blotting and the densities of phosphorylated Akt and EGFR (arbitrary units) are indicated above the blots. C, HeLa cells were transfected with scrambled, PKCα, and/or DGKδ siRNA duplexes, grown in medium with serum, and then the levels of the indicated proteins in cell lysates were detected by Western blotting and the densities of phosphorylated Akt and USP8 (arbitrary units) are indicated above the blots.