Fig. 2

Analysis of gene expression in overexpression mutants. a Quantification of reporter fluorescence by flow cytometry. Yellow fluorescence levels, corresponding to tdTomato fluorescence emission, were increased in all mutant strains. Fluorescence levels of NoGPAT-M1 and M2 were higher than for AoGPAT mutants, which suggests stronger expression of the endogenous GPAT compared to the heterologous gene. TdTomato fluorescence of NoGPAT-M2 was substantially decreased compared to NoGPAT-M1, likely due to the partial tdTomato gene sequence deletion found for this mutant (Additional file 1: Fig. S1b). b Western blot analysis of GPAT gene expression for NoGPAT-M1 and AoGPAT-M1. Protein extract was separated by SDS-PAGE, and recombinant GPAT was detected by immunoblotting for two biological replicates (R1 and R2) of NoGPAT-M1 and AoGPAT-M1 with 1 min and 15 min exposure times during chemiluminescence detection, on a single membrane. In line with the results of flow cytometry analysis, chemiluminescence signals were substantially higher for NoGPAT-M1 compared to AoGPAT-M1, suggesting more efficient expression of the endogenous gene compared to the heterologous one. The RuBisCO large subunit band of the Coomassie-stained membrane (C) is shown as a loading control