Inhibition of translation inhibits CHMP4B accumulation at the midbody. (A) HeLa cells were either untreated or treated with puromycin for 1 h and subjected to immunostaining with anti-acetylated-tubulin and anti-CHMP4B antibodies. The asterisks mark the MB and the white dashed square represents the region of the image used for the inset. Scale bars: 10 μm. Inset scale bars: 2 μm. (B and C) Shown data represent CHMP4B intensity/μm² in the intercellular bridge of 60 cells from three separate experiments. B shows distributions derived from individual cells (each dot represents a single cell). Statistical analysis in C was done on the means and SD derived from the three individual experiments where experimental means were calculated by averaging values from all the cells from each experiment. Each experiment is represented by a different shade of color. Statistical analysis is represented with a P value. (D and E) Shown data represent CHMP4B intensity/μm² in the cell body of 60 cells from three separate experiments. D shows distributions derived from individual cells (each dot represents a single cell). Statistical analysis in E was done on the means, and SD were derived from the three individual experiments where experimental means were calculated by averaging values from all the cells from each experiment. Each experiment is represented by a different shade of color. Statistical analysis is represented with a P value. (F) HeLa cells were either untreated or treated with puromycin for 1 h and subjected to immunostaining with anti-acetylated-tubulin and anti-CEP55 antibodies. The asterisks mark the MB and the white dashed square represents the region of the image used for the inset. Scale bars: 10 μm. Inset scale bars: 2 μm. (G and H) Shown data represents CEP55 intensity/μm² in the intercellular bridge of 60 cells from three separate experiments. G shows distributions derived from individual cells (each dot represents a single cell). Statistical analysis in H was done on the means and SD were derived from the three individual experiments where experimental means were calculated by averaging values from all the cells from each experiment. Each experiment is represented by a different shade of color. Statistical analysis is represented with a P value. (I and J) Shown data represents CEP55 intensity/μm² in the cell body of 60 cells from three separate experiments. I show the distributions derived from individual cells (each dot represents a single cell). Statistical analysis in J was done on the means, and SD were derived from the three individual experiments where experimental means were calculated by averaging values from all the cells from each experiment. Each experiment is represented by a different shade of color. Statistical analysis is represented with a P value. (K) HeLa cells after puromycin treatment were fixed and stained with anti-acetylated-tubulin antibodies. The percentage of cells in telophase was then counted. Shown data are the means and SD derived from three independent experiments. Statistical analysis is represented with a P value. (L) HeLa cells after puromycin treatment were fixed and stained with anti-acetylated-tubulin antibodies. The percentage of cells that just completed abscission was then counted. Shown data are the means and SD derived from three independent experiments. Statistical analysis is represented with a P value.