FIGURE 8.

TORC2 regulates dissociation of eIF4E·4EBP-1 complex and formation of eIF4E·eIF4G complex. 293 cells were transfected with either shRictor 1 (A and E), shRictor 2 (B and F), mSin1 sh1 (C and G), or mSin1 sh2 (D and H) expression vectors. Forty-eight hours post-transfection, the cells were lysed, and the extracts were immunoprecipitated with 4EBP-1 followed by immunoblotting (IB) with eIF4E and 4EBP-1 antibodies (A–D). E–H, the same cell lysates were immunoprecipitated (IP) with eIF4E followed by immunoblotting with eIF4G and eIF4E antibodies. Phosphorylation of Akt at Ser-473 to demonstrate TORC2 activity in rictor-down-regulated cells and expression of rictor and mSin1 in the cell lysates are shown in the bottom panels. Scr, scrambled; p-Akt, phospho-Akt.