Fig. 2. Mitochondria-targeted Kv1.3 inhibitors induce tyrosine phosphorylation of PI-3-K, AKT and JNK. Jurkat cells were left untreated or treated with 0.25 mM, 1 mM or 10 mM PCARBTP in the presence or absence of the anti-CD3 antibody OKT-3 (1 mg/mL) for the indicated times. Cells were lysed and proteins were separated by SDS-PAGE as above. Blots were developed for phospho-PI-3-K (p85) (a), phospho-AKT (b) or phospho-JNK (c). The results show a strong constitutive phosphorylation/activation of PI-3-K, AKT and JNK upon treatment with low doses of PCARBTP, which is only slightly further increased by OKT-3 stimulation.. Panels in Fig. 2. were performed from the same aliquots as in Fig. 1a and the corresponding actin blot is displayed in Fig. 1a. Blots were again quantified using Image J, which is displayed in the bar graphs in each Fig... Displayed are representative blots from 4 independent studies or the mean ± SD, n=4, *p<0.05, **p<0.01, ***p<0.001, ANOVA.