FIG 2.

Laccase is crucial for intracellular proliferation and survival of C. gattii within macrophages. (A) Laccase activity of LAC1 deletion strains (lac1Δ) was determined and compared with that of the parental (R265) and LAC1 complementary (lac1Δ::LAC1 [::LAC1]) strains. (B to I) The J774 macrophage was infected by C. gattii R265, lac1Δ, and complementary strains at a 1:10 ratio, followed by washing out the extracellular yeasts after 2 h of infection and then lysis at 2, 18, and 24 h postinfection to determine cryptococcal uptake (B) and phagocytic index (C) at 2 h, intracellular loads (ICL) at 18 h (D) and 24 h (E), intracellular proliferation rate (IPR) at 18 h (F) and 24 h (G), and percent killing at 18 h (H) and 24 h (I) using the CFU assay. Graphs depict means ± SD and are representative of three experiments, and significance was determined by one-way ANOVA with Tukey’s post hoc analysis; *, P < 0.05; **, P < 0.01; ***, P < 0.0001.