Fig. 4

NAMPT genetic ablation induces EIF2A phosphorylation and MCL1 down-regulation. a Expression levels of NAMPT. Jurkat cells were treated with 5 and 100 nM FK866 for 48 h. One representative experiment out of three biological replicates is presented. b WB analysis indicated 50 % of NAMPT silencing (p < 0.001) in Jurkat cells by using lentiviral particles expressing two NAMPT-silencing shRNAs (shNAMPT-1 and −2). c Intracellular NAD⁺(H) and ATP levels in shNAMPT cells (transduced with shNAMPT-1 and −2) were evaluated in comparison with scramble Jurkat cells. Thirty percent reduction of NAD⁺(H) levels was observed in shNAMPT cells (p-value < 0.01). RLUs were normalized to number of viable cells. Mean and SD of a biological triplicate. d WB analysis of AMPK, p-AMPK, EIF2A, p-EIF2A, p-4EBP1 and MCL1 in shNAMPT (transduced with shNAMPT-1 and −2) cells. Histogram shows the densitometric analysis of p-AMPK, p-EIF2A and MCL1 (* indicates p-value <0.05). Mean and SD of a biological triplicate