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. 2012 Oct 26;34(2):257–267. doi: 10.1093/carcin/bgs337

Fig. 3.

Fig. 3.

PrLZ induces the alteration of AR conformation and promotes nuclear translocation of AR (A–G). AR conformation alteration was analyzed by limited trypsin digestion in the presence of 10nM DHT or activator PrLZ (A). Results showed new different AR digested peptides (labeled as #, PrLZ + AR) or increased intensity of AR digested peptides (labeled as *, 10nM DHT + AR) as compared with control (labeled as $, AR). AR translocation was analyzed by immunofluorescence staining of AR. Cells were serum starved for 24h prior to performing the experiment. Data showed that PrLZ promotes AR translocation from cytosol to nuclei in PrLZ stably transfected LNCaP cells (LNCaP-PrLZ) (B), and in PC3-AR9-PrLZ (D) cells compared with vector control-transfected cells. In contrast, AR nuclear staining was significantly reduced in C4-2 cells transfected with PrLZ-siRNA (F) compared with scramble RNA-transfected control cells. 4′,6-Diamidino-2-phenylindole DNA staining indicates the locations of cell nuclei. Merged images of 4′,6-diamidino-2-phenylindole and AR IHC staining photos showed increased AR nuclear localization. Magnification ×400. The quantitation of PrLZ promoted AR nuclear translocation was further determined by assaying AR protein levels in nuclear versus cytoplasmic fractions in LNCaP (C), PC3-AR9 (E) and C4-2 cells (G).