FIG 3.

RAS1 and EFG1 deletions improve survival of C. albicans within human neutrophils. (A) H-PMNs were infected with C. albicans WT, Δ/Δras1, ras1/RAS1, RAS1OE, Δ/Δcyr1, and Δ/Δefg1 strains for 30 min. Extracellular fungal cells were stained using calcofluor white, unstained Candida cells were counted, and the PI was calculated by obtaining the ratio of the total number of ingested C. albicans cells and the total number of phagocytes counted. Phagocytosis of the WT was set as 100%. (B) Survival of C. albicans strains within H-PMNs was evaluated after 3 h of coincubation. H-PMNs were lysed, and internalized C. albicans cells were released, plated in agar, and incubated for 48 h to obtain viable CFU. Survival was calculated as follows: (recovered C. albicans CFU after neutrophil lysis/total number of phagocytosed C. albicans cells) × 100. Results represent the means ± SEs from at least three independent experiments carried out in duplicate. Significance was calculated using one-way ANOVA with post hoc Dunnett’s multiple-comparison test. *, P < 0.05; ****, P < 0.0001.