Figure 5

Characteristics of the CPAR2_200390Δ/Δ strain. (A) Growth of wild-type (CLIB 214), CPAR2_200390Δ/Δ and CPAR2_200390 reintegrant (RI) strains after 2 days on YPD solid media at selected temperatures. (B) Colony morphology on YPD plate following 2 days of incubation at 30 °C. (C) Pseudohyphae formation in 10% FBS supplemented DMEM medium after 24 hours of incubation at 37 °C. (D) Growth on 60 μg/ml calcofluor white supplemented YPD medium at 30 °C. (E) Fluorescence microscopic images of calcofluor white, ConA (Concanavalin A) and WGA (wheat germ agglutinin) stained wild-type, CPAR2_200390Δ/Δ and CPAR2_200390 RI cells. (F) Localization of chitin oligomer accumulations in the wild-type and mutant cell wall. (G) Relative normalized expression of C. albicans chitinase and chitin synthase orthologs (CPAR2_805640 as CHS1; CPAR2_701490 as CHS2; CPAR2_801800 as CHS3; CPAR2_502940 as CHS8, CPAR2_800050 as CHT1; CPAR2_ 502140 as CHT2, CPAR2_ 200660 as CHT3 and CPAR2_211950 as CHT4) after cultivation in YPD and 10% FBS supplemented DMEM medium. Statistical significance was determined by unpaired two-tailed t-tests (*p ≤ 0.05; **p ≤ 0.01; ****p ≤ 0.0001). Virulence attributes of the CPAR2_200390Δ/Δ strain following (H) an in vitro killing assay with J774.1 macrophage-like cells and after (I) G. mellonella larvae and (J) BALB/c mice infection. Statistical significance was determined by two-way ANOVA and Dunnett’s multiple comparisons tests in case of J774.1, and unpaired Mann-Whitney test in case of BALB/c mice infection. Mantel-Cox (Log-rank) tests were applied in the case of G. mellonella infection (*p ≤ 0.05; ****p ≤ 0.0001).