Effects of panobinostat (LBH589) and everolimus (RAD001) combination on the PI3K/AKT/mTOR pathway in HL cell lines. (A) Model showing PI3K/AKT/mTOR signaling. Everolimus inhibits mTOR/RAPTOR and leads to a down-regulation of p70S6K and eiF4E. (B-C) Cells were incubated with diluent control, LBH589 (0.02-0.1μM), RAD001 (0.1μM), or the combination of LBH589 and RAD001; and after 48 hours, cell lysates were prepared and resolved by SDS-PAGE, transferred to nitrocellulose membrane, and probed with (B) anti–phospho-AKT (Ser473), anti-AKT, anti–phospho-LKB1, anti-LKB1, anti–phospho-AMPKα, anti-AMPKα, anti–phospho-TSC2, anti-TSC2 antibodies, and (C) anti–phospho-RAPTOR, anti-RAPTOR, anti–phospho-mTOR, anti-mTOR, anti–phospho-p70S6K, anti-p70S6K, anti–phospho-4EBP1, anti-4EBP1, and anti–β-actin antibodies. Representative blots from 3 independents experiments are shown.