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Fig. 4. Activation of AR by agonistic ligands results in intranuclear localization in foci. Confocal laser-scanning microscope images showing representative nuclei of Hep3B cell lines stably expressing GFP-AR (A-C), the non-DNA-binding mutant GFP-AR(A573D) (D-F) or GFP-AR proteins with mutations in the LBD (which result in altered ligand specificity), GFP-AR(W741C) (G-I) and GFP-AR(T877A) (J-L). Subnuclear localization was observed in the presence of 10–9 M R1881 (A,D,G,J), 10–6 M OH-flutamide (B,E,H,K) or 10–6 M bicalutamide (C,F,I,L). With all ligands, androgen receptors are localized in the nucleus, but are excluded from nucleoli (dark areas in the nucleus). In situations where AR is able to activate transcription intranuclear foci are observed (A,G,I,J,K, see Fig. 1). Bar, 5 µm.





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