Figure 2. Protein production inside artificial cell-like particles.

Confocal microscopy images of sfGFP production inside protein producing liposomal particles after 60 minutes (A), and as a function of time (B), both at 37°C. sfGFP was imaged in the green channel, and the rhodamine-labeled liposome membrane was imaged in the red channel. (C) 3D image of the protein producing particles after 2 hours. (D) A Western blot analysis demonstrates that sfGFP (~27kDa) is produced both in bulk and in the particles. (E) Flow cytometry of sfGFP producing liposomes: Hoechst was used for nuclei staining, rhodamine was incorporated in the particle' membrane, and sfGFP was produced in the particles. The particles were analyzed for sfGFP synthesis (F), as a function of particle diameter (µm) (G). (H) sfGFP production kinetics inside artificial cell-like particles and in a cell-free bulk. A linear regression was fitted to the first third of each of the reactions incubation times. Error bars represent standard deviation of the mean from 2 independent repeats. (I)&(J) Artificial cell-like particles source molecular building blocks from their surrounding environment to trigger protein production. (I) An illustration of adding molecular building blocks that are necessary for transcription and translation through the particle membrane. (J) The effect of amino acids and nucleotide supplementation on sfGFP production inside particles. The fluorescence was measured after a 4 hour incubation. Error bars represent standard deviation of the mean from at least 3 independent repeats. Treatments that differ significantly by a two-tailed Student’s t test (p-value<0.05) are designated with different letters (a, b & c).