Fig. 4: Stoichiometry optimization of the three components for the influenza A subtype H1N1 VLP via the predictive model. | Nature Communications

Fig. 4: Stoichiometry optimization of the three components for the influenza A subtype H1N1 VLP via the predictive model.

From: Precise programming of multigene expression stoichiometry in mammalian cells by a modular and programmable transcriptional system

Fig. 4

a Schematic of the recombinant VLP in which NA and M1 proteins are fused with mCherry and GFP reporter proteins, respectively. b Transmission electron microscopy images of influenza VLPs. Scale bar, 50 nm. c Representative images of the VLPs produced from the three-gene system. Fluorescent microscopy was used to observe co-localization events of NA and M1. Scale bar, 10 μm. d Schematic of the plasmid constructions for VLP production. e Relative VLP yields for 21 selected promoter-gene combinations and 3 widely-used mammalian promoters (EF1α, CMV and SV40). The reference VLP yield was of the combination of the strongest promoters and was assigned as 100%. Data represent the mean ± SD (n = 3). f Predicted VLP yields in all 157,464 combinations of the available HA, M1 and NA promoters. The reference combination is normalized to 1. The dashed circle marks the rough boundary of the promoter combinations with yields higher than that of the reference. Stars indicate the combinations selected for experimental validation. g Comparison of experimental VLP yields with model predictions. Data represent the mean ± SD (n = 3). Source data are provided as a Source data file.

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