Effects of the inhibitors on class I PI3K/Akt/mTOR axis signaling in canine C2 cells. Cells were treated with pan-class I PI3K inhibitor Wortmannin (W) at 1 μM and ZSTK474 (Z) at 1 μM, mTOR inhibitor Rapamycin (R) at 100 nM (A) and Akt inhibitor KP372-1 at 0, 150, 200 and 400 nM (B) for the indicated period of time. 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. N/A indicates data unavailable due to induction of apoptosis in all cells.