Metformin inhibits translation and decreases the expression of oncogenic proteins in AML. (A) 7-mGTP affinity assays of primary AML samples cultured for 24 hours without or with 5, 10, or 15mM metformin (samples 14 and 19 are representative of 5 independent experiments). (B) Polysome analysis performed in the MV4-11 cell line cultured for 12 hours, without or with 10mM metformin. (C) Results of GeneChip (Affymetrix) analysis of RNA extracts from the sucrose gradients used to extract polysomes. The diagram represents mRNA whose polysome recruitment was decreased (green lines) or increased (red lines) in control (CTR1 and CTR2) and metformin (MET1 and MET2) replicates. (D) [³H]leucine pulse assays performed in primary AML samples treated for 2 hours without or with 5, 10, or 15mM metformin. Ratios to the control incubation without an inhibitor were then calculated, and the results are presented as a mean of 5 independent experiments. Vertical bars indicate the SEM. (E) Quantification of Western blot signals. Ratios of c-Myc, cyclin D1, or Bcl-xL to actin signal intensity were calculated, and results are expressed relative to the control conditions (without metformin) in each experiment. Each histogram represents the mean of 5 independent experiments using primary AML samples. The vertical bars indicate the SEM.