Fig. 3

Visualization and quantification of mitochondrial distribution in dividing HeLa cells expressing KIF5BTail and Miro-1ΔTM. a Spinning disk confocal images of HeLa cells transfected with control vector (top two rows), KIF5BTail (middle two rows) or Miro-1ΔTM (bottom two rows) and stained with MitoTracker Deep Red FM to visualize mitochondria. Images are single z slices from the centre of the confocal stack and maximum z-stack projections are shown for five representative time points from metaphase to late-cytokinesis. Blue arrowheads indicate mitochondria that are mislocalized are the cell poles in KIF5BTail- and Miro-1ΔTM-expressing cells. Bar, 10 μm. b Quantification of mitochondrial fluorescence intensity from cell pole to equator in control (14 cells, N = 56), KIF5BTail- (14 cells, N = 56), and Miro-1ΔTM-expressing cells (16 cells, N = 64) during metaphase, anaphase, and early-, mid- and late-cytokinesis. The normalized distance from cell pole to equator is displayed on the x-axis and the average fluorescence intensity normalized against the mean is displayed on the y-axis. Data are represented as the mean ± SEM and lines fitted by non-linear regression. The difference in mitochondrial distribution was statistically significant in late cytokinesis in KIF5BTail-expressing cells (F-Test, p < 0.05) and from early cytokinesis onwards in Miro-1ΔTM-expressing cells (F-Test, p < 0.05) compared with control cells