NaChl inhibits phosphorylation of Abl in K562 cells in a time- and dose-dependent manner without affecting Src-family kinases and JAK kinases. (A) K562 cells were treated with varying concentrations of NaChl for 30 minutes (left panel) or with 10 μg/mL NaChl for varying time periods (right panel), fixed, permeabilized, stained with anti–phospho-c-Abl (solid line) or normal rabbit sera (filled histogram), and analyzed in a flow cytometer. (B) NaChl treatment reduces Abl-protein expression in a time-dependent manner in K562 cells. Cells were treated with NaChl (10 μg/mL) for indicated times, fixed, permeabilized, and stained with anti–c-Abl antibody (□, cultured in medium; ○, cultured in the presence of NaChl). The level of expression is presented as mean fluorescence intensity (MFI). Data represent mean ± SD of 3 experiments. (C) Immunoblot-based determination of protein expression and phosphorylation status of indicated kinases in indicated cells before (NT) and after treatment for 30 minutes with NaChl (T). Experiments were performed as mentioned in the legend of Figure 3A using indicated antibodies.