FIGURE 2.

mTORC1/mTORC2 inhibition decreases the phosphorylation of downstream targets. C57BL/6 albino mice were implanted with KPC pancreatic cancer cells and injected intraperitoneally daily with Torin 2 at a high dose, Torin 2 at a low dose, metformin at a low dose, both Torin 2 and metformin at low doses (TLML), or DMSO control for 12 d. Mice were killed, and the pancreas was collected, flash-frozen in liquid nitrogen, and stored at –80°C until analysis. Whole-cell lysates were harvested and immunoblotted with total mTOR, P-mTOR Ser2481, total Akt, P-Akt-Ser473, total S6, P-S6, or β-actin antibody. C, control; ML, metformin at low dose; mTORC, mechanistic target of rapamycin complex; TH, Torin 2 at high dose; TL, Torin 2 at low dose; TLML, Torin 2 and metformin at low doses.