Supplemental Material

This article contains the following supporting material:

• Figure 1 - Subcellular fractionation of cells transfected with hFis1, hFis1^K148R or Control Plasmid
  MEFs transfected either with the indicated plasmids and after 24 hrs mitochondrial, cytosol and light membranes (LM) fractions were purified as described in experimental procedures. Equal amounts (40 μg) of protein were separated by SDS-PAGE and immunoblotted using the indicated antibodies.

• Figure 2 - Mitochondrial movement is not impaired by hFis1.
  MEFs were co-transfected with mtRFP and β-gal or hFis1 and after 24 hours mtRFP was imaged by confocal microscopy. Individual frames were acquired every 5 s over a 2 min time-frame. In representative images acquired at the indicated times single moving mitochondria were pseudocolored in green for the sake of clarity. Red bars show point of origin of moving mitochondria. Bar, 1 μm.

• Figure 3 - Staurosporine induces mitochondrial fragmentation but not apoptosis in DKO MEFs
  (A) Apoptosis was determined by flow cytometry as the percentage of Annexin-V, PI positive cells. Data represent mean ± SE of 5 independent experiments. Where indicated, STS (1 μM) was added 10 hrs before assay. (B) Representative images of mitochondrial morphology in wt and DKO MEFs transfected with mtRFP. 24 hrs after transfection STS (1μM) was added where indicated and fluorescence of mtRFP was visualized by confocal microscopy 8 hours later. Bar, 15 μm