Fig. 2

ITK inhibition reduced proliferation and inhibited TCR signaling in malignant T cells. a Jurkat, Hut-78, and H9 cells were transfected with vectors carrying ITK shRNA (shITK-34465, shITK-34466 and shITK-34467) or a nonspecific shRNA (shControl). Viable cells were analyzed using the Cell Titer-Glo Luminescent Cell Viability Assay. b Jurkat, Hut-78, and H9 cells transfected with shITK or shControl were subjected to Western blotting analysis of p-PLCγ1, p-Akt, Akt, p-Erk, and Erk. GAPDH is shown as a loading control. c Karpas-299 cells were transfected with carrying ITK shRNA (shITK-34465, shITK-34466 and shITK-34467) or a nonspecific shRNA (shControl). Viable cells were calculated using the Cell Titer-Glo Luminescent Cell Viability Assay. d Jurkat, Hut-78, H9, Karpas-299 and T cells from healthy donors were treated with the indicated concentrations of the ITK inhibitor BMS-509744. Viable cells (% control) were calculated. e Jurkat, Hut-78, and H9 cells were pretreated with BMS-509744 (3 μM, 5 μM, or 8 μM) for the indicated times (60 min, or 90 min), and the whole-cell extracts were subjected to Western blot. f Jurkat, Hut-78, and H9 cells were pretreated with BMS-509744 (3 μM, 5 μM, or 8 μM) for 24 h, and the whole-cell extracts were subjected to Western blot. Data are expressed as Mean ± SD and representative of three independent experiments. Data are expressed as Mean ± SD and representative of three independent experiments