Figure 5.

Androgen promotes YAP1 nuclear localization. A and B, coimmunofluorescence imaging of YAP1 and AR proteins in LNCaP cells. The cells were treated with or without DHT (10 nm) and ENZ (20 μm) overnight in 5% CSS-fed growth conditions. Images of AR (Alexa 647, red), YAP1 (Alexa 488, green), and nuclei (DAPI, blue) were acquired using confocal microscopy. Scale bar, 20 μm. The micrographs are the representation of multiple images. B, ImageJ software was used to quantify the intensity of the nuclear AR and YAP1 protein from multiple images. The data ± S.D. normalized to the total cell number from multiple images of three independent experiments. *,**, p < 0.0001. C, coimmunoprecipitation and Western blotting analysis of the cytoplasmic and nuclear YAP1, PP2A C, and AR proteins in LNCaP cells after treatment with or without DHT overnight in 5% CSS-fed growth condition. Topoisomerase I (TOPI) was used as a nuclear marker. The blots are representative of three independent experiments. MW, molecular mass.