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. Author manuscript; available in PMC: 2017 Nov 1.
Published in final edited form as: Mol Cancer Res. 2016 Aug 26;14(11):1054–1067. doi: 10.1158/1541-7786.MCR-16-0167

Figure 3. AR and ER co-localize in the nucleus in response to E2.

Figure 3

(a) MCF7 cells were grown in media with CSS for 72 hr then pre-treated with veh, 10 uM Enza, or 1 uM bicalutamide (bic). Following pre-treatment, cells were treated with veh or 10nM E2 +/− Enza or bic as shown for an additional 3 hr. Cells were then fixed and ICC was performed for AR (green) and ER (red). (b,c) MCF7 cells were grown in media with CSS for 72hrs then treated with the indicated treatment for 3 hrs, and nuclear extracts were immunoblotted for AR and TOPO1. (d) ER+/AR+ ZR-75-1 or (e) ER−/AR+ MDA-453 cells were grown in media with CSS for 72hrs, then pre-treated for 3 hr with Enza or vehicle control. Following pre-treatment, cells were treated with veh, 10nM DHT, or 10nM E2 +/− Enza as shown for 3 additional hrs. Nuclear extracts were then obtained and subjected to western blotting for AR and TopoI. (f) MCF7 cells were grown in media with CSS for 72hrs then treated with E2 +/− Enza for 1 h followed by fixation and PLA staining for AR and ER (red). Nuclei were stained with DAPI (blue). (g) Fluorescent intensity per nuclei was measured by CellProfiler. Error bars represent standard error of the mean. ****p <0.0001 by ANOVA with Dunnett’s Multiple Comparison Test.