PLK4OE and MCAK depletion, respectively, induce centrosome amplification and increased astral MT length. (A) Pictures of Ctrl (DOX-) and PLK4OE (DOX+) OVCAR-8 cells labeled with antibodies against CEP192 and Centrin 3 to label the centrosomes and against the membrane marker N-cadherin. (B) Graph bars showing the percentage of Ctrl (DOX-; n = 488 cells from two independent experiments) and PLK4OE (DOX+; n = 410 cells from two independent experiments) OVCAR-8 cells presenting four or fewer or more than four centrioles. Statistical significance was determined using a multiple t test. (C) Western blot of siCtrl and siMCAK, OVCAR-8 cells. (D) Pictures of siCtrl polyploid and siMCAK polyploid OVCAR-8 cells labeled with antibodies against α-tubulin. (E) Dot plot showing the length of astral MTs in siCtrl, polyploid (n = 140 MTs from 11 cells) and siMCAK, polyploid (n = 118 MTs from eight cells) OVCAR-8 cells. Statistical significance was determined using a Mann-Whitney test. (F) Stills of time-lapse video of mitotic polyploid OVCAR-8 cell with stabilized MTs (siMCAK) expressing histone 2B-GFP (red and gray in the middle insets) and incubated with SiR-tubulin (green and fire LUT in the bottom insets). Orange and white dotted circles surround cells and nuclei, respectively. Time of mitosis is represented in minutes:seconds, and time 00:00 corresponds to the NEBD. Error bars represent the mean ± SD. p corresponds to the P value.