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Figure 2


Fig. 2. Expression of caveolin-1 in HEK293T cells promoted cell death. HEK293T cells were transfected with pLacIOP or pLacIOP-caveolin-1 as indicated and analyzed after 24 hours. (A) Viability was determined by the Trypan Blue exclusion assay. (B) To determine apoptosis, cells were harvested, resuspended in PBS with 10 µg/ml propidium iodide (PI) and analyzed by flow cytometry. In A and B values were averaged from three independent experiments (mean ± s.e.m.). #comparison with pLacIOP control (P<0.05), §comparison with pLacIOP control (P<0.1). (C) Results from a standard DNA laddering assay. From left to right, samples from non-transfected (lane 1), pLacIOP-transfected (lane 2) and pLacIOP-caveolin-1-transfected (lane 3) cells are shown. (D) Nuclear fragmentation analysis. Caveolin-1 was detected by immunofluorescence (green) in cells transfected with pLacIOP-caveolin-1 and the integrity of the nuclei was assessed by PI staining (red). Samples were analyzed by confocal microscopy (Bar, 10 µm). In C and D a representative result from two independent experiments is shown.





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