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. 2022 Dec 20;51(1):99–116. doi: 10.1093/nar/gkac1158

Figure 1.

Figure 1.

AR foci form upon androgen stimulation. (A) Foci formed by transiently expressed and endogenous AR (Endo AR). Cells were cultured with 5% CSS-containing medium for two days then stimulated with 1 nM DHT or ethanol (control) for 2 h. The transiently expressed AR-mEGFP and the immune-stained endogenous AR were inspected under confocal microscope. (B) Time-lapsed foci formation. LNCaP cells expressing AR-mEGFP were hormone starved (5% CSS) for 2 days and then stimulated with 1 nM DHT. A live time-lapse confocal imaging assay was performed along the DHT treatment. (C) Reversibility of foci formation. AR-mEGFP expressing LNCaP cells were starved in 5% CSS for 2 days, treated with DHT for 2 h, and then washed twice to remove DHT. Cells were then cultured in 5% CSS for the indicated time course. DHT was applied back to cells at 6h and 24 h post-DHT removal, respectively. AR-rich foci were quantified. Percentage of AR-mEGFP foci containing cells were quantified from 45 fields of three independent experiments and the data was presented as aligned points with mean ± SD. (D) Androgen dependence of foci formation. AR-mEGFP expressing LNCaP cells were cultured in 5% CSS for 2 days then treated with various hormones and foci formation was quantified as in 1C. One-way ANOVA was used to analyse the data in (C) and (D). P values are indicated by stars: ns ≥ 0.05, * 0.01 to 0.05, ** 0.001 to 0.01, *** 0.0001 to 0.001, **** < 0.0001.