Figure S1 - The Combination of RAD001 and NVP-BEZ235 Exerts Synergistic Anticancer Activity against Non-Small Cell Lung Cancer In Vitro and In Vivo

Concurrent combination of rapamycin and LY294002 is more effective than sequential treatments in inhibiting the formation and growth of NSCLC colonies. The indicated cell lines at a density of approximately 200 cells/well were seeded in 24-well plates. On the second day, cells were treated with 1 nM rapamycin (Rap) alone, 5 nM LY294002 (LY) alone, concurrent combination of rapamycin and LY294002 (Rap+LY), rapamycin for 3 days and then switched to LY294002 treatment (Rap→LY), LY294002 for 3 days and then switched to rapamycin treatment (LY→Rap). The same cycles of the treatments were repeated every 3 days. After 12 days, the plates were stained for the formation of cell colonies with crystal violet dye. The picture of the colonies was then taken using a digital camera (A) and the colonies were counted (B). (PDF)