Longer polyQ sequences increase the tendency of AR to localize to nuclear puncta in the presence of DHT.

(A) Representative images of cells stably expressing Cherry-AR-12Q, -20Q, -31Q or -49Q, treated with DHT 24 h before fixation. Red corresponds to Cherry fluorescence and blue to DAPI fluorescence. (B) Puncta intensity ratios (IRs) in cells stably expressing Cherry-AR-12Q, -20Q, -31Q or -49Q were calculated as the ratio of fluorescent intensity in the puncta minus the background divided by fluorescent intensity in the nucleoplasm minus the background. The background intensity corresponds to an image taken from an area where there were no cells. Bars represent average IRs calculate from measurements performed in at least 50 cells under each condition (5–45 puncta per cell) and errors bars show standard deviations. Statistical significance and P values were determined by one-way analysis of variance (ANOVA) with Holm-Sidak test (** P < 0.01; *** P < 0.001).