Figure 6: TFs involved in the pathogenicity of C. neoformans.

(a) Novel virulence-regulating TFs identified using the G. mellonella killing assay. P values shown in the graph were calculated using the Log-rank test to measure statistical differences between the WT strain (H99) and each TF mutant strain. The insect infected with a second independent mutant for each TF exhibited similar survival patterns (Supplementary Fig. 8). (b) Virulence-regulating TFs identified using the signature-tagged mutagenesis (STM)-based mouse study. The y axis indicates the average STM scores from two independent mutants for each TF. The STM score for each mutant was calculated using quantitative PCR performed using a common primer and the signature-tag-specific primers listed in Supplementary Data 2. The ste50Δ and ire1Δ mutants were used as virulent and non-virulent control strains, respectively, based on previous reports^9,55. Blue-dotted lines indicate cutoffs (±1.5) that we set to define meaningful changes in virulence based on the STM scores of the ste50Δ mutant. Mutants illustrated in red are reported for the first time in this study. Virulence-related TFs were divided into two groups. In the high-confidence group (marked as an asterisk (*)), TF mutants exhibit high average STM scores (>2 or <−2) and each STM score of two independent mutants is statistically significant (P<0.05 compared with ste50Δ mutant) and beyond the cutoff. In the remaining low confidence group, the average STM scores for TF mutants range from −2.0 to −1.5 or 1.5 to 2.0 with significant P values (<0.05), but one of STM scores from two independent mutants could be within the cutoff, although both STM scores have a similar trend (both positive or negative values). The P value between control and mutant strains was calculated by one-way analysis of variance with Bonferroni's multiple-comparison test with three biological replicates (three mice per set of experiments). Error bars indicate s.e.m.