SNX9 depletion phenocopies CHC depletion during mitosis.

<p>HeLa cells were either untreated (-), or transfected with luciferase siRNA (Luc) or siRNA targeting CHC (A), SNX9 (B), SNX18(C) and SNX33 (D). At 72 h post-transfection, protein lysates (200µg) were immunoblotted with anti-CHC, anti-SNX9, anti-SNX18 and anti-SNX33 antibodies respectively. Actin was used as a loading control. (E–G) HeLa cells were treated with siRNA targeting Luc, CHC, SNX9, SNX18 and SNX33 and visualised by time-lapse microscopy. Graphs show the time that each individual cell took to undergo mitosis (n > 100 per sample; E) as well as the following mitotic transitions: prophase to anaphase (F), and anaphase to completion (G). Pro, prophase; Ana, anaphase; Comp, completion. Data shown in E-G are from one representative experiment. Similar results were obtained in at least two independent experiments. The median time per sample is represented by a solid black line. ***, <i>p</i> <0.001 (One-way ANOVA).