Figure 4. Phenotypic diversity of the flipped progeny recovered from infected mice.

A, A quantitative representation of phenotypic assays of the flipped and the non-flipped progeny recovered from animal lungs and brains after animals were euthanized at DPI 13. Cells were diluted into OD₆₀₀=0.03 and spotted onto YPD agar media in a 96-spot format with the indicated stress factors at the indicated concentrations. Images were taken after four days of incubation at 30°C and analyzed using Colonyzer [51]. The median growth index values are shown as black bars. See Figure S4 and Table S1&S2 for the raw resource images, data, and the strain list. Mann-Whitney test was used to analyze the growth difference between WT, the flipped, and the non-flipped strains under zeocin condition. The p values of the pairwise comparisons were shown. B, Serial dilutions of the selected flipped and non-flipped progeny were spotted onto YPD agar media supplemented with the indicated stress factors at the indicated doses. Cells were then incubated at 30°C for four days. The parental haploid H99 strain and a diploid strain (H99α/H99α) served as controls. The strain labels correspond to their plate positions in Figure S4B. C, A time course growth assay of the flipped, non-flipped, and WT strains cultured in liquid YPD with or without zeocin. The overnight cultures of the strains used in panel B were diluted into OD=0.05 and inoculated into liquid YPD or YPD with 10 μg/ml of zeocin in a 48-well plate. The plates were cultured at 30°C with continuous shaking and growth was monitored by measuring OD₆₀₀ every hour. The growth of the WT, the non-flipped, and the flipped strains based on OD₆₀₀ measurement was plotted against the time after inoculation (growth curve assay). The colored range shows the distribution of growth curves of the WT (black), the non-flipped (orange), and the flipped (green) strains. See also Figure S4.