Figure 4

Effects of the inhibitors on class I PI3K/Akt/mTOR axis signaling in canine and human cancer cells. Cells were seeded at a density of 20,000 cells per ml overnight, followed by treatment with 1 μM ZSTK474 (A), 400 nM KP372-1 (B), or 100 nM Rapamycin (C) for 5 hrs. Whole cell lysates (comprising 50 μg total protein) were subjected to western blot with the indicated antibodies. β-actin was used as a loading control.