Fig. 3

PLK4 and AURKB/C redundantly modulate ACM-induced cancer cell motility. a MDA-MB-231 cells were treated with DMSO or different concentrations of CFI-400945 (PLK4 and AURKB/C inhibitor), in the presence of DMEM or ACM (****p < 0.0001; n = 3, at least 50 cells were tracked per condition). b MDA-MB-231 cells were treated with DMSO or 100 nM CFI-400945 in the presence of ACM stimulation. Cells were stained for α-tubulin and actin, and enlarged images showed a cortical region of the cell. Bar = 20 µm (n = 3). c MDA-MB-231 cells were treated with centrinone, centrinone B (PLK4 inhibitors), AZD (AURKB/C inhibitor) or their combination, in the presence of DMEM or ACM overnight (***p < 0.0001; n = 3, at least 50 cells were tracked per condition). d Cells from (c) were stained for α-tubulin and actin, and cortical regions were enlarged on right side panels. Bar = 20 µm. e Cells were incubated with DMEM or ACM overnight. At 6 h of taking time-lapse images, DMSO or inhibitors were added to cells and incubation continued for 14 h more, then DMSO or inhibitors were washed out and cells were further incubated in DMEM or ACM for another 16 h. Running average speed was plotted as mean ± s.e.m. (n = 4, at least 50 cells were tracked per condition) f Breast cancer cell lines EpRas and MDA-MB-468 were treated with DMEM or ACM in the presence of DMSO or inhibitors (**p < 0.01, ****p < 0.0001; n = 3, at least 40 cells were tracked per condition). All the data were plotted as box and whiskers. Boxes represent median and 25th to 75th percentiles, whiskers the minimum and maximum values with each individual cell value superimposed. The data were compared with one-way ANOVA Kruskal–Wallis test and post-tested with Dunn’s multiple comparison test. Arrow indicates the control bar used for comparison