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. Author manuscript; available in PMC: 2022 Aug 18.
Published in final edited form as: J Am Chem Soc. 2021 Aug 4;143(32):12675–12687. doi: 10.1021/jacs.1c05386

Figure 1|. Characterization of NBDY-RNA coacervates.

Figure 1|

(a) TMR-NBDY or polyU RNA alone does not form phase separated liquid droplets. Transmission brightfield (left) and fluorescence images (right). (b,c) Transmission brightfield (left) and fluorescence images (right) show coacervate phase droplets formed by 0.4% w/v polyU RNA (polyuridylic acid) and 200 μM TMR-NBDY (b) or by 0.5 μg/μL HEK 293T RNA and 200 μM TMR-NBDY (c). (d) 0.5 μg/μL FITC-labelled U40 RNA and 200 μM TMR-NBDY colocalize in coacervate droplets. Buffer: 25 mM HEPES, pH 7.4, 0.5 mM sodium azide, 0.5% w/v Tryptone, 10% PEG3350 at 30 °C. Scale bars, 10 μm. (e,f) Turbidity plot of NBDY-polyU coacervates at increasing concentration of NBDY (e) or NaCl (f). 0.4% w/v polyU was used in all cases, and 200 μM NBDY was used in (f). Error bars in turbidity plots represent the standard deviation of three independent trials. Buffer: 25 mM HEPES, pH 7.4, 0.5 mM sodium azide, 0.5% w/v Tryptone, 10% PEG3350 at 30 °C.