Mitochondrial puncta containing both Caf4 and Mdv1 are fission competent.

<p>(<b>A</b>) Large panel at the left shows a representative image of <i>caf4</i>Δ <i>mdv1</i>Δ cells expressing GFP-Caf4 and RFP-Mdv1. Scale bar: 5 μm. a-f marks puncta enlarged from the image in (A) at the left. Examples of partially or completely co-localized (a–c) versus isolated (d–f) puncta are shown. Scale bar: 0.2 μm. (<b>B</b>) Quantification of Caf4 and Mdv1 colocalization. GFP- or RFP-tagged Caf4 and Mdv1 were expressed in <i>caf4Δ mdv1Δ</i> cells in the pair-wise combinations indicated. The number of GFP puncta colocalized with RFP puncta was quantified and normalized to the total number of RFP puncta in each cell (n = 10 cells). Bars and error bars are the mean and SD of three independent experiments. (<b>C</b>) Time lapse imaging of a mitochondrial fission event at a site where Caf4 and Mdv1 co-localize. Cerulean-tagged Caf4 (shown in cyan pseudo-color) and EYFP-Mdv1 were integrated at the <i>HO</i> and <i>MDV1</i> loci, respectively. Mitochondria were visualized using plasmid-borne mt-mCherry. The overlay contains true color images of all three channels. White areas indicate regions where signals from all three fluorescent proteins overlap. Scale Bar: 0.6 μm.