Brf1 is critical for the protective effect elicited by ZNF2 overexpression. (A) Phenotypical analyses of H99, ZNF2oe, brf1Δ, and ZNF2oebrf1Δ on thermo- or CO2- tolerance (YPD at 30°C, 30°C + 10% CO2, and 37°C) and melanization (L-Dopa medium at 22°C). (B) The lung fungal burden at day 12 postinoculation with live H99 (5 mice), ZNF2oe (5 mice), brf1Δ (5 mice), or ZNF2oebrf1Δ (5 mice) at the inoculum of 1 × 104 cells/animal. (C) Animal survival was monitored for 60 days after inoculation with live H99 (10 mice) or ZNF2oebrf1Δ (10 mice) at 1 × 104 cells/animal. The survival rate was plotted against the days postinoculation. (D) The lung fungal burden of the surviving mice inoculated with live ZNF2oebrf1Δ (10 mice) at the time of termination (DPI 60) compared the lung fungal burden of mice infected with live H99 (10 mice) at time of euthanization when they reached defined clinical endpoints. (E) Survival of mice vaccinated with live ZNF2oebrf1Δ (10 mice) or ZNF2oe (10 mice) after challenge with live H99. (F) Survival of mice vaccinated with heat-killed ZNF2oebrf1Δ (10 mice) or ZNF2oe (10 mice) after challenge with live H99.