Fig. 2

miR-221-5p exerts tumor suppressive function on PCa cell lines in vitro. a Left: Relative miR-221-5p expression (2^-ΔCt) in normal prostatic epithelial Ep156T cells and AR⁺ and AR⁻ PCa cell lines. Analysis by one-way ANOVA with Tukey’s multiple comparisons test. Right: Technical replicates of at least two independent experiments for each cell line. miR-221-5p expression of AR⁺ cells was analysed by one-way ANOVA with Tukey’s multiple comparisons test. AR⁻ cell lines were compared by unpaired, two-tailed t-test. * p < 0.05, *** p < 0.001, **** p < 0.0001. Figure symbols: cross: Ep156T; circle: LNCaP; square: C4–2; triangle: VCaP; filled circle: PC-3M-Pro4luc2; filled square: DU145 cells. b miR-221-5p expression levels in PC-3M-Pro4luc2 cells upon overexpression or knock-down of miR-221-5p. LOG difference to control was calculated as LOG(2^-ΔΔCt) and analysed by unpaired, two-tailed t-test. Three technical replicates of representative experiments are shown. **** p < 0.0001 vs. scrambled, ## p < 0.01 vs. anti-scrambled. c Effect of miR-221-5p on proliferation. Images were taken 72 h post transfection with miR-221-5p and scrambled. MTS assay at 0 h, 24 h, 48 h, 72 h. Fold change compared to time point T’0 h was calculated for each time point. Data of n = 4 independent experiments are shown and were analysed by two-way ANOVA repeated measures by both factors with Sidak’s multiple comparison test. * p < 0.05, ** p < 0.01. d Effect of anti-miR-221-5p on proliferation. Images were taken at 72 h post transfection. MTS assay at 0 h, 24 h, 48 h, 72 h. Fold change compared to time point T’0 h was calculated for each time point. Data represent n = 5 technical replicates and were analysed by two-way ANOVA repeated measures with Sidak’s multiple comparison test. e Clonogenicity assay in miR-221-5p overexpressing PC-3M-Pro4luc2 cells. Dots indicate technical replicates of n = 4 independent experiments. Fold change was calculated vs. scrambled control and analysed by unpaired, two-tailed t-test. *** p < 0.001. f Clonogenicity assay in miR-221-5p knocked-down PC-3M-Pro4luc2 cells. N = 4 technical replicates are shown as fold change to control and analysed by unpaired, two-tailed t-test