Phagocyte recruitment to hypovirulent C. albicans mutants. (A) Example representative images from brg1∆/∆, pep8∆/∆ and cek1∆/∆-infected fish, along with SN250-infected controls, at 4–6 hours postinfection. Images were scored by eye for the number of macrophages (mpeg1:GFP+ cells) shown in green and the number of neutrophils (lysC:dsRed+ cells) in magenta recruited to the infection, as well as if the Candida was intracellular or extracellular. Scale bar is 100 µm. (B–D) Quantification of phagocyte recruitment-related phenotypes. There are separate SN250 columns for each set of experiments, as the mutant was compared to wild type in the same experiments. (B) Plots showing the number of mpeg:GFP+ macrophages recruited to the infection site normalized to the average amount of mpeg:GFP+ macrophages recruited to SN250. (C) Plots showing the number of lysC:dsRed+ neutrophils recruited to the infection site normalized to the average amount of lysC:dsRed+ neutrophils recruited to SN250. (D) Plots showing the number of cells recruited to the infection site normalized to the average recruited to SN250. Cells include mpeg1:GFP+ and lysC:dsRed+ cells recruited to the hindbrain, as well as non-fluorescent cells containing Candida. (B–D) Shading indicates Groups I–IV based on similar interaction phenotypes (Fig. 4). Means and 95% confidence intervals are plotted. Statistics were performed from data pooled from at least three independent experiments for each mutant, for approximately 25 fish per strain were imaged. Hedges bias-corrected effect sizes and significance were determined for each mutant. * indicates P < 0.05, # indicates a moderate effect, while a bold # indicates a strong effect.
|
