FIGURE 6.

Loss of hnRNP K leads to caspase-dependent and caspase-independent cell death. HeLa cells were transfected with control or hnRNP K siRNA in the presence or absence of zVAD-fmk (100 μm). A, 3 days post-transfection, cells (3,750 cells/well) were seeded in 96-well plates and 5 days post-transfection, activity of caspase-3 and -7 was assessed by subjecting cell lysates to Caspase-Glo 3/7 Assay. Bars represent the mean ± S.D. of triplicates. B, cells (1 × 10⁵ cells/well) were seeded in 6-well plates 3 days post-transfection, and apoptosis was assessed by staining cells with annexin-V-FLUOS and PI followed by flow cytometry analysis 5 days post-transfection. Background apoptotic values of control siRNA-treated cells were subtracted and the apoptotic value of hnRNP K siRNA-treated cells without zVAD-fmk was set to 100%. Bars represent the mean with range of two individual experiments. C, 3 days post-transfection, cells were seeded in E-plates at different cell densities (3,750 or 7,500 cells/well), and overall cell behavior was monitored real-time using xCELLigence. Bars represent the mean ± S.D. of at least two individual experiments performed in triplicate. *, p < 0.01; **, p < 0.001.