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Figure 6


Fig. 6. Mitochondrial fragmentation-stimulating activity of MIB is offset by the coexpression of Mfn proteins. (A) Mfn1-HA, MIB-FLAG, or both were transfected to HeLa cells and the mitochondrial structures of the cells were examined by immunofluorescence microscopy as described in Materials and Methods. Bars, 10 µm. (B) The cells containing fragmented mitochondria were quantified. At least 100 cells were counted in three distinct optical fields. (C) Mfn1-FLAG or Mfn2-FLAG (Eura et al., 2003) was coexpressed with MIB-HA in HeLa cells. The mitochondria were isolated from the cells and solubilized by 1% digitonin. The solubilized fractions were subjected to immunoprecipitation using anti-HA IgG under the indicated conditions. The immunoprecipitates were analyzed by SDS-PAGE and subsequent immunoblotting with anti-HA or anti-FLAG IgGs.





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