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. Author manuscript; available in PMC: 2020 Jul 2.
Published in final edited form as: Biochemistry. 2018 Apr 19;57(18):2590–2596. doi: 10.1021/acs.biochem.8b00131

Fig. 4. Triggering formation of membraneless organelles in cell-like compartments using small molecule and light-inducible protease activity.

Fig. 4.

(A) Schematic of fluorescently-tagged IDP formation of membraneless organelles. MBP solubilization domain is cleaved from the IDP, resulting in phase separation and formation of membraneless organelles. (B) Rapamycin-dependent formation of protein droplets within emulsions. In absence of rapamycin, IDP remains soluble and well mixed. 1 μM split TEV was +/− equimolar dimerizer and 30 μM IDP, in the presence of 25% Xenopus egg extract. (C) Light-induced TEV activation and formation of membraneless organelles in cell-like compartments. Compartments encapsulated with 1 μM split TEV, 500 nM dRap, 30 μM IDP, 25% egg extract; either kept in dark or exposed to 365 nm UV light for 10 minutes. For 4B-C, emulsions were imaged 12 hours post-induction. Scalebar: 20 μm.