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Fig. 5. Overexpression of hFis1 compromises steady state mitochondrial morphology. COS-7 cells overexpressing the GFP-hFis1 chimera were counterstained with MitoTracker Red and Hoechst 33258 and analysed by fluorescence microscopy. Cells expressing hFis1 mainly displayed one of two phenotypes: (A) fragmented mitochondria; or (B) mitochondrial aggregates in the vicinity of the nucleus. (C) Phenotype of cells expressing the control outer membrane protein Tom7. (D) High-resolution confocal microscopy of mitochondrial aggregates similar to those seen in (B) reveals that they are composed of small fragmented mitochondria. (E) Quantitation of mitochondrial aggregates in COS-7 cells overexpressing vector alone, Tom7 or hFis1 was monitored and quantified at 16 hours, 36 hours and 48 hours after transfection (n=3). (F) Quantitation of mitochondrial aggregates in >100 COS-7 cells co-expressing hFis1/Drp1 or hFis1/Drp1 (K38A) (n=3).





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