Figure 4

Influence of Taxol treatment on cell viability: (a)Three-color fluorescence viability assay (top and middle) and hematoxylin/eosin staining (bottom) of slices treated with different taxol concentrations for 72 hours (images of a representative experiment). (b) The numbers of TMRM⁺, SYTO^®63⁺, and Picogreen⁺ cells in tumor slices from (a) were evaluated by counting of at least 50 cells from three different areas of different images. Viable cells were assessed as numbers of TMRM⁺ and SYTO^®63⁺ cells in relation to total cells (left panel). Dead cells were evaluated as numbers of Picogreen⁺ cells in relation to total cells (right panel). Values reflect means ± SEM. (c) Dose dependent effect of taxol treatment on cell viability determined by quantification of TMRM⁺, SYTO^®63⁺, and Picogreen⁺ cells in non fixed tumor slices (left and middle panel) and by evaluation of ATP/DNA ratio in slice homogenates (right panel). Ratios of controls were set to 100%, and the relative ratios of the taxol treated tissues were calculated. Data represent range, median and 25 to 75% percentile of a set of 10 experiments. Kruskal-Wallis test: p < 0.0001 for all three. (d) Effect of taxol treatment on proliferation and apoptosis. Formalin-fixed and paraffin-embedded slices were stained with anti-BrdU or anti-caspase 3 antibodies by conventional immunohistochemistry and counterstained with hematoxylin.