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Fig. 7. Cytochrome c release and cell death are delayed in cells overexpressing Myc-hFis1[32-152]. Myc-hFis1[32-152] or Myc-hFis1-FL was co-expressed with GFP as a transfection marker and distributions of cytochrome c were analyzed by immunofluorescence. (A) Cytochrome c staining was greatly reduced and diffuse in cells expressing Myc-hFis1-WT, indicating cytochrome c release from mitochondria (a,a'). Cells expressing Myc-hFis1[32-152] contained cytochrome c in their swollen mitochondria (b,b'). T: transfected cells. Scale bar, 10 µm. (B) Cells with diffuse cytochrome c staining were scored at different times after transfection. Less than 25% of cells transfected with Myc-hFis1[32-152] released cytochrome c up to 72 hours post transfection, whereas 70% of cells expressing full-length hFis1 released cytochrome c at 72 hours. (C) Frequencies of cells expressing full-length hFis1, hFis1[32-152], or both Bcl-2 and hFis1[32-152] were scored at 24, 48 and 72 hours post transfection and presented as percent cell survival at each time point. The frequency at 24 hours was given as initial survival (100%). Survival of cells expressing full-length hFis1 decreased to 20% after 72 hours, whereas that of hFis1[32-152] cells were 75-80% at the same time point. Co-expression of Bcl-2 with Myc-hFis1[32-152] increased cell survival to 95% after 72 hours.
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