Fig. 5: LC coacervate microdroplets with integrated biomolecules reconfigured into isotropic yolk-shell coacervate vesicles.

a Scheme showed the transformation of LC coacervate microdroplets with integrated proteins or nucleic acids into organelle-containing coacervate vesicles induced by the hydrolysis of Cm-Am. Time-lapse fluorescence microscopy images displaying LC microdroplets with the incorporation of RITC-BSA (0.3 mg/mL) (b) or TAMRA-ssDNA (99 nt, 1.0 μM) (c) reconfigured into yolk-shell coacervate vesicles initiated by amylase (0.9 mg/mL). Time sequence of fluorescence microscopy images exhibited homogenous distribution of RITC-Cm-Am during the reconfiguration of HRP-integrated LC microdroplets (d), while the LC microdroplets with integrated FITC-dextran (10 k Da, 0.5 mg/mL) transformed into coacervate vesicles in the presence of amylase (0.9 mg/mL) (e). f Time-dependent FRAP kinetics as evaluated by fluorescent probe of FITC-PEI for LC microdroplets with integrated HRP, ssDNA (99 nt), or dextran (10 k Da), LC microdroplets without biomolecules, and rim or inner organelle within yolk-shell coacervate vesicles. The data represented mean ± s.d. (n = 3 independent experiments). g Reconstructed 3D merged fluorescence microscopy image displaying an artificial organelle-containing coacervate vesicle was formed after reconfiguration of LC droplets with the simultaneous integration of Py-HRP, TAMRA-ssDNA (99 nt), and FITC-dextran (10 k Da). It exhibited nearly white fluorescence, indicating the homogenous distributions of these biomolecules. Grid width = 1 µm. Schematic illustration of HH-min-mediated cleavage of a FRET-ssRNA substrate to liberate the fluorophore from the quencher strand (h) and the corresponding ribozyme reaction was supported within artificial organelle-containing coacervate vesicles (i). Time-dependent fluorescence microscopy images of a yolk-shell coacervate vesicle (j), Cm-Am/DDAB LC microdroplet (k), and the corresponding fluorescence intensity plots (l) exhibited faster reaction rates within rim and inner organelle than that in the LC coacervate microdroplet. The data represented mean ± s.d. (n = 3 independent experiments). Scale bars, b–e, 5 μm; j, k, 2 µm. The experiments for b–g, j–k were repeated three times independently with similar results. Source data are provided as a Source data file.