Fig. 6: Condensate pump-mediated biological processes.

a Schematic illustration of DNA unwinding and green fluorescent protein (GFP) synthesis controlled by condensate pumps (Azo Coas500) for transmembrane transporting DNase and plasmid GFP DNA. In the DNA unwinding process, the amount of DNase_31kDa pumped into proteinosome was adjusted by the number of pumping cycles (1 or 3 times). During the GFP synthesis, proteinosomes encapsulating E. coli lysate containing a large amount of transcription and translation-related factors and Q-Dex_500kDa were co-incubated with Azo Coas500 loaded with plasmid GFP DNA. GFP DNA was pumped into proteinosomes, thereby constructing a cell-free protein expression system within protocell communities. In- situ CLSM images of DNA unwinding after once pumping (b), thrice pumping (c) of Azo Coas500. DNA is labeled with SYBR Green I (SYBR Green-DNA, green fluorescence); RBITC-DNase, red fluorescence. Scale bars, 25 μm. d Fluorescence intensity changes of DNA unwinding induced by DNase for once pumping (circles, blue line) and thrice pumping (squares, green line) of Azo Coas500. Control group (rhombuses, gray line) shows fluorescence quenching of SYBR Green-DNA without DNase. e Fluorescence intensity at different positions of proteinosomes after GFP synthesis. Box plots show the mean line (midline), 25/75 percentiles (boxes), and SD (whiskers). Data are represented as mean ± SD (n = 5). Positions 1~4 were shown in (f) and 5 was shown in Supplementary Fig. 35. f CLSM image of GFP synthesis within proteinosomes. Scale bar, 3 μm. All relevant experiments in b, c are performed independently at least three times with similar results. Source data is provided as a Source Data file.