Fig. 1. A mutagenesis screen uncovered T. gondii mutants exhibiting decreased GFP expression driven by the LDH2 promoter.

a Illustration of ENU-mediated mutagenesis and FACS for enriching parasite mutants with increased autophagy signal during the bradyzoites stage. b Representative flow cytometry pseudocolor plots demonstrating the intensities of tdTomato-ATG8 and GFP (driven by the bradyzoite-specific LDH2 promoter) for parasites before or after three rounds of sorting. The final enriched population (pop’n) (red dashed box) were subcloned and subjected to whole genome sequencing. c Quantification of GFP and tdTomato-ATG8 intensities before and after three rounds of sorting. As different voltage settings were used for GFP and tdTomato-ATG8 for before and after the three rounds of sorting, direct comparison of absolute median fluorescence intensity values is not feasible. Therefore, median fluorescence intensities for GFP or tdTomato-ATG8 in the mutant population were divided by those in the WT to obtain the ratio of median fluorescence intensity (mut pop’n/WT), as depicted in the bar graphs. The numbers in the pseudocolor plots represent the percentage of the gated population relative to the total population. Source data are provided as a Source Data file.