Figure 5: Aurora A regulates AR-V expression.

(a) Co-immunoprecipitation was carried out using the AR (N-20) antibody on HEK293 cells overexpressing FL-AR, AR-V7 or AR-123/2b and Aurora A followed by subsequent western analysis using anti-AR (N-20) and –Aurora A antibodies. Input samples were also subjected to western blot analysis (bottom panel). (b) Impact of Aurora A depletion on AR levels was measured by western blot analysis of CWR22Rv1 cells transiently transfected with Aurora A-targeting siRNA. (c) LNCaP cells were transiently transfected with siAURKA and incubated for 6 hours prior to transduction with lentivirus expressing AR-V7 for 72 hours. Resultant lysates were subjected to western blot analysis. (d) Parallel samples of those described in (b) were subjected to quantitative-PCR to measure AR-V7 and FL-AR mRNA expression following Aurora A depletion. Data is represented as fold-change of siScr. Data represents three independent experiments ± SEM. P-values were determined by one-way ANOVA followed by a Bonferroni multiple comparison test (****denotes p-value < 0.001). (e) CWR22Rv1 cells were transiently transfected with siAURKA or siScr for 72 hours with the addition of 20 μM MG132 (MG) for the final 4 or 8 hours before harvest. AR levels were measured by western blot analysis using the AR (N-20) antibody.