Fig. 4.
(a) Relative expression of CDH1 and VIM mRNA in MCF7_miR200c_KO and DOX-treated MDA-MB-231_i-miR200c (MB231_i-miR200c) cells normalized to wild type (unmodified MCF7 or untreated MB231_i-miR220c, respectively). (b) Western blot of MB231_i-miR200c cells with and without doxycycline-induction stained for EMT-relevant marker CDH1, VIM and CD44s. (c) Whiskers plot showing alterations in CN-values (from 40× images) in MB231_i-miR200c cells with and without doxycycline-induction for growth on TACS5- and TACS6-like structures. One-way ANOVA with Bonferroni multiple comparison test was performed to calculate P-values at a 95 % confidence interval (n (cells) > 50). (d) Corresponding confocal images of (c) at 10× magnification. Cells were stained for nuclei (blue) and the cytoskeleton (red). Comparison of trajectory analysis of (e) WT cells with MCF7_KO or (f) MB231_i-miR200c, respectively, performed with Fiji software (TrackMate). Displacement, mean migration speed, confinement ration and mean directional change were plotted as whiskers plot (5–95 percentiles). Statistical significance was assessed with a t-test based on > 150 cells/condition. Stars indicate statistical significance (∗∗∗P < 0.001; ∗∗P < 0.01; ∗P < 0.05, ns = not significant).