Figure 10

ID2 and ID3 gene knockdown reduces survival of MEC1 cell line and primary CLL cells. (A) Western analysis of MEC1 cells infected with lentiviruses encoding either a control siRNA sequence or encoding one of four siRNAs targeting different sequences in the ID2/ID3 mRNAs. The original western blot images are shown in Additional file 10: Figure S5. (B) MEC1 cells that were transduced with control siRNA (open bars), ID2R-siRNA (light-shaded bars) or ID3Y-siRNA (dark-shaded bars) were incubated in the absence or presence of increasing concentrations chlorambucil or ethacrynic acid for 48 hrs and cell viability was assessed by MTT assay. The mean ± SEM of three independent samples is shown. (C) CLL cells from four patients were transfected with 60 nM negative control siRNA or with ID2/ID3 siRNA. 72 hrs post-transfection, cells were harvested and analysed by western blotting. The original western blot images are shown in Additional file 10: Figure S5. (D) 1×10⁶ cells from the four CLL patients were transfected with 60 nM negative control siRNA (open bars) or with the same concentration of ID2 siRNA (light-shaded bars) or ID3 siRNA (dark-shaded bars). 48 hrs post-transfection, the cells were incubated in the absence or presence of fludarabine (5 μM) for a further 24 hrs. Cell viability was assessed by MTT assay and values were normalised to the untreated control siRNA sample. Data from the mean ± SEM from three independent samples is shown.