Fig 7

(A) Susceptibility of WT CEA10, ΔzfpA, and OE::zfpA to 0.25, 0.5, 1, and 8 μg/ml caspofungin (CSP). 10⁴ spores were point-inoculated on solid GMM with caspofungin or DMSO. Bars represent mean±s.d. of colony diameter at 4 days post inoculation of 4 plates per condition. (B) Susceptibility of WT CEA10, ΔzfpA, and OE::zfpA to 0.25, 0.5, 1, and 8 μg/ml micafungin (MCF). 10⁴ spores were point-inoculated on solid GMM with micafungin or DMSO. Bars represent mean±s.d. of colony diameter at 4 days post inoculation of 4 plates per condition. p values calculated by ANOVA with Tukey’s multiple comparisons. (C) Images represent calcofluor white (CFW) staining of WT CEA10, ΔzfpA, and OE::zfpA following overnight exposure to 1 μg/mL caspofungin (CSP) or DMSO. CFW staining is represented by cyan and cytoplasmic RFP signal is shown in magenta. Scale bar = 50 μm. (D) Mean gray value of CFW signal following DMSO or caspofungin treatment. Bars represent mean±s.e.m of 3 independent experiments. n = 24–30 hyphae per condition. (E) Experimental setup for chitin stimulation with CaCl₂/CFW. Spores were incubated for 8 h at 37°C or until germination in liquid GMM or liquid GMM supplemented with 0.2 M CaCl₂ and 100 μg/mL CFW. After germination, media was replaced for GMM + 1 μg/mL caspofungin or DMSO and hyphae were incubated for an additional 12 h before detecting PrestoBlue viability reagent signal in a plate reader. (F) Bars represent mean±s.d. of relative fungal viability following caspofungin exposure. Relative viability was calculated by normalizing the mean signal of caspofungin-treated wells to the mean signal of DMSO-treated wells. All experiments included 5 wells/condition. Data are pooled from 3 independent experiments. p values calculated by ANOVA with Sidak’s multiple comparisons. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.