Fig. 2.

Asymmetric partitioning of mitochondria during mitosis and stochastic failure of cytokinesis in Myo19-KO cells can be rescued by Myo19, but not by individual Myo19 domains. (A) Immunoblot verifying the generation of Myo19-deficient cell lines stably expressing different Myo19 constructs as indicated on top. Immunoblots were developed with antibodies directed against Halo-Tag, β-actin and VDAC1, respectively. Arrowheads point at the respective constructs. The dashed lines reflect the borders of a membrane slice. (B) Cellular localization of Halo-tagged constructs. Cells were transfected with Mito-EGFP to label the mitochondria and incubated with TMR-ligand to label the indicated Halo-fusion constructs (red). Scale bar: 20 µm. (C) Cell proliferation was analysed for different cell clones as indicated; mean±s.e.m., n=4 independent experiments. (D) Quantification of the number of nuclei per cell in the different cell clones as indicated; n=3 independent experiments, N≥159 cells. (E) Quantification of DNA content by flow cytometry analysis of cells stained with propidium iodide (PI). Data are represented as mean±s.e.m. for 4 independent experiments. (F) Quantification of mitochondria distribution at cell poles versus spindle at anaphase for cells of the indicated cell lines. (G,H) Quantification of the asymmetric mitochondria distribution in cells at anaphase and telophase. Data in F–H are displayed as box plots with the sample median of at least 20 cells, together with the corresponding 75th and 25th percentiles. The whiskers show the 10th–90th percentiles. The minimum and maximum values are highlighted with dashes. n≥7. **P≤0.01; *P≤0.05; n.s., not significant (one-way ANOVA with Bonferroni post-hoc test and Mann–Whitney U-test).