Additional file 2: of EIF2A-dependent translational arrest protects leukemia cells from the energetic stress induced by NAMPT inhibition

Luciferase assays. A) Light units, normalized to protein concentration, of RLuc-cMyc 5′UTR IRES-FLuc reporter vector transduced in Jurkat cells with lentiviral particles after 48 h of treatment with or without (Mock) the indicated concentration of FK866. Two hour treatment with 250 nM of Torin 1 served as a positive control for IRES-dependent protein translation (p-value <0.05). B) Light units, normalized to protein concentration, of FLuc-HCV-RLuc and FLuc-CrPV-RLuc reporter vectors transduced in Jurkat cells with lentiviral particles. Cap-dependent translation (FLuc) was strongly reduced with 5 nM and 100 nM FK866 (48 h) in comparison to Mock condition (p-value <0.0001). RLuc signal is not shown because of its low level and its variability between technical and biological replicates. Cells transduced with the pHR-SIN-F-HCV-R were serum starved for 5 h as a positive control of IRES activation, as shown in the graph (p-value <0.05). In A and B data are represented as mean and SD of three independent experiments. (PDF 584 kb)