Fig. 1: Reprogramming the PKR-dependent eIF2α phosphorylation pathway enables GCE enhancement in mammalian cells.

a General scheme of GCE. RS designates aminoacyl-tRNA synthetase, ncAA - noncanonical amino acid. b Schematic representation of the PKR-dependent eIF2α phosphorylation pathway. c Schematic representation of the iRFP-GFP^39TAG reporter. The gray population represents untransfected cells; gray arrow – CMV promoter; gray box – polyA signal, FC denotes flow cytometry. d Heat map illustrating the effect of adding various stress remodelers on GCE efficiency in units of relative efficiency^GFP (%). Percentage relative efficiency^GFP is calculated as the median GFP signal for each particular case divided by the median GFP signal for control samples with the addition of mock plasmid. Median GFP signals were obtained after FC analysis of corresponding samples. The heat map shows the mean value for the relative efficiencies of three independent experiments. Source data are provided as a Source Data file. e FC analysis of the iRFP-GFP^39TAG reporter in the absence and presence of the tested stress remodelers. Concatenated data from three independent experiments (50 ng tRNA^Pyl plasmid) are shown.