Effect of SFK inhibitors on phosphorylation of survival mediators in CLL. (A) Freshly isolated CLL cells were cultured in the absence or presence of 0.1 µM dasatinib, 10 µM PP2, or 1µM saracatinib for 24 hours, and cell apoptosis was analyzed by annexin V–PI flow cytometry. Quadrant analysis after flow cytometry was expressed as mean percentage ± standard deviation (SD) of early and late apoptosis from 3 separate experiments performed in triplicate on samples from 30 CLL patients. Compared with the control, changes were statistically significant (*P < .01). (B) Freshly isolated CLL cells were cultured in the absence or presence of 0.1 µM dasatinib, 10 µM PP2, or 1 µM saracatinib at different time points. After such treatment, cells were lysed and analyzed by western blot (Wb) analysis with anti-PARP antibodies, as well as anti-β-actin antibody as a loading control. (C) Freshly isolated CLL cells were cultured in the absence or presence of 0.1 µM dasatinib, 10 µM PP2, or 1 µM saracatinib at different time points. After such treatment, cells were lysed and analyzed by Wb analysis with antibodies against the phosphorylated form and, after stripping, against the nonphosphorylated form of Lyn, Akt, GSK-3, and SHP-1.