Figure 7: Inhibition of Aurora A reduces AR-V7 levels, reduces AR target gene expression and reduces proliferation in CWR22Rv1 cells.

(a–d) CWR22Rv1 cells were grown in steroid-depleted conditions and treated with 1 or 3 μM Aurora A inhibitor (S1451) for 24 hours. AR protein levels (a) and transcripts (b) were measured by respective western blot and quantitative PCR analyses. qPCR data is represented as fold-change of Vehicle (Veh). Data represents three independent experiments ± SEM. P-values were determined by one-way ANOVA followed by a Bonferroni multiple comparison test (*and **denote p-values < 0.05 and <0.01, respectively). (c) ASF/SF2 levels were measured by western blot analysis following Aurora A inhibition. (d) PSA, TMPRSS2 and CCNA2 expression was measured by quantitative-PCR. qPCR data is represented as fold-change of Vehicle (Veh). Data represents three independent experiments ± SEM. P-values were determined by one-way ANOVA followed by a Bonferroni multiple comparison test (*and **denote p-values < 0.05 and <0.01, respectively). (e) Effects of Aurora A inhibition of CWR22Rv1 cell proliferation was measured by subjecting CWR22Rv1 cells grown in steroid-depleted conditions to increasing doses of the Aurora A inhibitor (0–10 μM). Proliferation was measured using the IncucyteZoom live cell imager and normalised to control. Data represents three independent experiments ± SEM. P-values were determined one-way ANOVA followed by a Bonferroni multiple comparison test (*, **, ***and ****denote p-values < 0.05, <0.01, <0.001 and <0.0001, respectively).