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

Figure 3.

Figure 3.

Recombinant AR protein forms droplets in vitro. (A) Effect of PEG-8000 concentration on His-AR-mEGFP (0.25 μM) droplet formation as characterized by confocal microscopy and turbidity assay after 20 min at room temperature. Droplet formation is associated with increased turbidity and light scattering that could be followed by optical density measurements at 340 nm. (B) Droplets as a function of time. Droplets are formed 15–20 min after incubation with 10% PEG-8000. (C) Droplets as function of His-AR-mEGFPconcentration. The number of droplets increased with the increasing concentrations of His-AR-mEGFP. Effect of 1,6-hexanediol (D) and salt concentration (E) on His-AR-mEGFP droplet formation. His-AR-mEGFP at 0.25 μM was incubated for 20 min in presence of 10% PEG-8000 and increasing concentration of 1,6-hexanediol or NaCl. (F) FRAP assay showing fluorescence recovery after photobleaching of His-AR-mEGFP droplet in comparison to an unbleached droplet and the relative fluorescence intensity of the bleached droplets (mean ± SD, N = 3). Scale bars: 5 μm. 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.