Fig. 3.

Overexpression of miR-221-5p affects plasticity of PCa cells and reduces extravasation in vivo. a E-CAD/N-CAD and E-CAD/VIM ratios were calculated from the relative mRNA expression and data of n = 3 independent experiments are shown as fold change to control. Data were analysed by paired, two-tailed t-test. b Expression of E-CAD and VIM protein in PC-3M-Pro4luc2 cells 72 h post transfection with miR-221-5p and scrambled. Bands were quantified and E-CAD/VIM ratio was calculated (normalised to β-actin). The data of n = 2 independent experiments are shown as fold change to scrambled and were analysed by paired, two-tailed t-test. ** p < 0.01. c Migration of miR-221-5p overexpressing PC-3M-Pro4luc2 cells. Dots represent technical replicates of n = 3 independent experiments. Data are shown as fold change to scrambled and were analysed by unpaired, two-tailed t-test. **** p < 0.0001. d Migration of PC-3M-Pro4luc2 cells with miR-221-5p knock-down. N = 3 technical replicates are shown. Data were normalised to control and analysed by unpaired, two-tailed t-test. e Phalloidin staining in PC-3M-Pro4luc2 cells overexpressing miR-221-5p or scrambled control. F-actin signal was normalised to number of nuclei. Data was analysed by unpaired, two-tailed t-test. F-actin = green; DAPI = blue. f Extravasation of miR-221-5p and scrambled transfected PC-3M-pro4_mCherry cells at the caudal hematopoietic tissue (CHT) of zebrafish embryos 4 days post injection (dpi). Tumor burden was quantified 1dpi and 4dpi. Green = vessels; red = PC-3M-pro4_mCherry cells. Data was analysed by unpaired, two-tailed t-test. ** p < 0.01