Translation is reduced by 4EGI-1 but not by RAD001 in AML. (A) [³H] leucine pulses were performed to determine global protein synthesis rates. Blast cells from 5 AML patients and from the MOLM-14 leukemic cell line were cultured for 2 hours in a leucine-deficient medium, without or with 10 nM RAD001, 25 μM LY294002, or 50 or 100 μM 4EGI-1, and then pulsed with [³H] leucine (1 μCi, 37 kBq) for 1 hour. The amount of radioactivity incorporated into macromolecules was determined by trichloroacetic acid precipitation. Results are presented as the ratio to the control incubation without inhibitor. Vertical bars represent SD. (B) Blast cells from 5 AML patients were cultured for 24 hours in αMEM containing 10% FCS, without or with 10 nM RAD001 or 50 or 100 μM 4EGI-1. Cell extracts were analyzed by Western blot to assess the expression of c-Myc, Cyclin D1, Bcl-xL, and actin. The signal intensity was quantified, and results are expressed as the ratio between c-Myc, Cyclin D1, or Bcl-xL and actin signals, relative to the control condition. Vertical bars represent SD.