Fig. 6

(a). The effect of DZNep/Daunoblastine combination on apoptosis. FACS analysis after double labelling of  Jurkat cells  with PI and Annexin V. Jurkat were treated with DZNep and Daunoblastine alone and in combination, compared to the control. The experiments were performed at least three times and the results were always similar. Insets, % of positive cells. UL = Upper Left (necrosis); UR = Upper Right (late apoptosis); LL = Lower Left (viable); LR = Lower Right (early apoptosis). Untreated cells, CTR; Dauno added for 48 h, DZNep 48 h; DZNep + Dauno added for 48 h. (a). The figure is representative of three different experiments that always gave similar results. The table shows the percentage of cells in the different quadrants. (b). FACS analysis after double labelling Jurkat with PI and Annexin V. Jurkat were treated with DZNep and Daunoblastine alone and in combination, compared to the control at 72 h. (c). Cells were then processed for the determination of the expression of caspase 3, caspase 9 and Bcl2 evaluated after blotting with specific antibodies described in Materials and Methods section. The house-keeping protein beta-tubulin was used as loading control. CTR, untreated; Dauno, DZNep and Dauno/DZNep treated cells as described before. Each point is representative of three different evaluations performed in at least three different experiments. Scan of the bands associated with caspase 3, caspase 9 and Bcl2 expression in Jurkat cells, normalized with the house-keeping protein, was performed with a dedicated software and the intensities of the bands were expressed as percentage protein expression (%, mean of three different experiments). Bars, s.e.’s