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

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posted on 2015-11-05, 05:00 authored by Chiara Zucal, Vito D’Agostino, Antonio Casini, Barbara Mantelli, Natthakan Thongon, Debora Soncini, Irene Caffa, Michele Cea, Alberto Ballestrero, Alessandro Quattrone, Stefano Indraccolo, Alessio Nencioni, Alessandro Provenzani
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)

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