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. 2018 Feb 28;293(17):6410–6433. doi: 10.1074/jbc.M117.813741

Figure 9.

Figure 9.

CgYPS-C genes are required for survival of the macrophage internal milieu and nitrosative stress. A, intracellular growth profiles of the indicated CgypsΔ mutants in THP-1 macrophages. THP-1 macrophage infection was done as described in the legend to Fig. 3A with C. glabrata cells at an MOI of 10:1. Fold Replication indicates the ratio of the number of intracellular C. glabrata cells at 24 h to that at 2 h postinfection. Data represent mean ± S.E. (error bars) of 3–5 independent experiments. **, p < 0.01; ****, p < 0.0001; unpaired two-tailed Student's t test. B, IL-1β was measured in the culture supernatant of uninfected THP-1 and THP-1 cells infected with the indicated C. glabrata strains as described in the legend to Fig. 6B. Data (mean ± S.E.; n = 3–5) represent secreted IL-1β levels under the indicated conditions. Statistically significant differences in IL-1β levels between uninfected and C. glabrata–infected and between WT– and Cgyps1–11Δ–infected macrophages are indicated by black and gray asterisks, respectively. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; unpaired two-tailed Student's t test. C, intracellular growth profiles of the Cgyps1–11Δ mutant overexpressing individual CgYPS1–11 genes in THP-1 macrophages. Fold Replication indicates the ratio of the number of intracellular C. glabrata cells at 24 h to that at 2 h postinfection. Data represent mean ± S.E. of 3–5 independent experiments. V, C. glabrata strains carrying empty vector. ***, p < 0.001; unpaired two-tailed Student's t test. D, IL-1β was measured in the culture supernatant of uninfected THP-1 and THP-1 cells infected with the indicated C. glabrata strains as described in the legend to Fig. 6B. Data (mean ± S.E.; n = 4) represent secreted IL-1β levels under the indicated conditions. V, C. glabrata strains carrying empty vector. Statistically significant differences in IL-1β levels between WT– and Cgyps1–11Δ strain–infected macrophages are marked. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; unpaired two-tailed Student's t test. E, intracellular survival of Cgyps1Δ and Cgyps1–11Δ mutant expressing either CgYps1 or putative catalytically dead CgYps1D91A in THP-1 macrophages. Fold Replication indicates the ratio of the number of intracellular C. glabrata cells at 24 h to that at 2 h postinfection. Data represent mean ± S.E. of 3–7 independent experiments. Black asterisks, statistically significant differences between WT and mutants carrying either vector or CgYps1D91A protein. Gray asterisks, statistically significant differences between mutants carrying vector and CgYps1 protein. **, p < 0.01; ****, p < 0.0001; unpaired two-tailed Student's t test. F, serial dilution spotting analysis of CgypsΔ mutants under the indicated conditions. Menadione, MMS, and sodium nitrite (NaNO2) were used at a final concentration of 50 mm, 0.04%, and 60 mm, respectively. Growth was recorded after 2 days of incubation at 30 °C.