DEDs impair tumor growth, proliferation and trigger defects in mitosis. A. Scheme of GFP-tagged caspase-8 derivatives, which express either the DEDs alone, the catalytic domain active or inactive (C360A), the holoenzyme (caspase-8B) active or inactive (C360A). B. Immunoblot analysis showing expression of GFP, caspase-8 inactive-GFP (C8*-GFP), caspase-8-GFP (C8-GFP), catalytic domain-GFP ... [Show full abstract] (CAT-GFP), catalytic domain inactive-GFP (CAT*-GFP) and DED-GFP. Upper panel: Western blotting with a C8 DEDs-specific antibody. Middle panel: Western blotting with a C8 Catalytic domain-specific antibody. Lower panel: Western blot with a tubulin-specific antibody, used as loading control. C. Proliferation assay performed with GFP, C8-GFP, C8*-GFP, CAT-GFP, CAT*-GFP and DED-GFP expressing cells. D. GFP, C8-GFP and DED-GFP expressing cells were stained with propidium iodide and percentage of apoptotic cells was measured by flow cytometry. Data were analyzed with U-Mann-Whitney Test (*, significant differences, p≤0.05; **, very significant differences, p≤0.01). E. Evaluation of tumor growth in chick embryos of NB7 neuroblastoma cells stably transfected with DED-GFP, Casp8-GFP, Casp8*-GFP or control GFP. F. Immunofluorescence images of human Small Cell Lung Carcinoma Cells (SCLC) transfected with DED-GFP or control GFP and Neuroblastoma NB7 cells transfected with DED-GFP, stained with lamin B (red channel) and a DNA dye (blue channel) show accumulation of micronuclei. White arrows show micronuclei or amorphous nuclei (scale bar = 10 µm).
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