Article Figures & Data
Figures
- FIG. 1.
GMS-stained (upper panels) and anti-GFP immunohistochemically stained (lower panels) tissue sections from C. albicans-infected mice. The strains used for inoculation are listed above each pair of panels. The left three panels (strains PTPI1-GFP, CAI12, and PALS1-GFP) show kidney tissue 48 h after infection. The right two panels (PALS1-GFP and PALS3-GFP) show heart tissue 24 h after infection. Inoculation of mice with the PTPI1-GFP strain produced strong, constitutive GFP production and obvious immunohistochemical staining. Staining of tissues infected with strain CAI12, which does not produce GFP, was performed as a negative control. Anti-GFP staining of strains producing GFP under the control of the ALS1 or ALS3 promoter was obvious in all kidney and heart sections in which GMS-stained fungal cells were present. Images were taken at ×400 magnification.
Tables
- TABLE 1.
Observations from GMS- and immunohistochemically stained C. albicans-inoculated mouse tissue sectionsa
Inoculation strain Time point (h) Resultb Kidney Heart Liver GMS Spleen GMS GMS IHC GMS IHC Uninfected 48 − − − − − − CAI12 12 +/+ −/− +/+ −/− −/+ −/− 24 +/+ −/− +/− −/− +/− −/− 48 +/+ −/− +/+ −/− +/+ −/− PTPII-GFP 12 +/+ +/+ −/− −/− −/+ −/+ 24 +/+ +/+ −/− −/− +/+ +/− 48 +/+ +/+ −/+ −/+ −/− −/+ PALS1-GFP 12 +/+ +/+ −/+ −/+ −/− −/− 24 +/+ +/+ +/+ +/+ +/+ −/+ 48 +/+ +/+ +/+ +/+ −/− −/− PALS3-GFP 12 +/+ +/+ +/+ +/+ −/− −/− 24 +/+ +/+ −/+ −/+ −/− −/− 48 +/+ +/+ −/− −/− −/− +/+ PALS5-GFP 12 +/+ −/− +/− −/− +/− +/− 24 +/+ −/− +/+ −/− +/− −/− 48 +/+ −/− −/+ −/− −/+ −/− PALS6-GFP 12 −/− −/− −/− −/− −/− −/− 24 +/+ −/− −/+ −/− −/− −/− 48 +/+ −/− −/− −/− −/− −/+ PALS7-GFP 12 +/+ −/− +/+ −/− −/+ −/− 24 +/+ −/− +/+ −/− +/− −/− 48 +/+ −/− +/+ −/− +/+ −/− PALS9-GFP 12 +/+ −/− −/− −/− −/− −/− 24 +/+ −/− −/− −/− −/− −/− 48 +/+ −/− −/− −/− +/− +/− ↵a Tissues from PALS-GFP-infected mice were analyzed using Gomori methenamine silver (GMS) staining and anti-GFP immunohistochemical (IHC) detection. Each entry in the table shows results from a pair of mice at each time point.
↵b +, presence of fungal element(s) stained black for GMS or brown for IHC visible at ×400 magnification. −, absence of fungal element(s) stained by GMS or lack of staining by IHC in the presence of GMS-positive fungal element(s). All lung sections were negative for GMS and IHC staining at all time points in all animals. All liver and spleen sections were negative by IHC staining, so only GMS results are reported in the table.
- TABLE 2.
Results of GFP-specific RT-PCRs from analysis of total kidney RNA from mice inoculated with C. albicans PALS-GFP reporter strains
Duration of infection (h) Inoculation straina PTPI1-GFP PALS1-GFP PALS3-GFP PALS9-GFP 12 +/+ +/− −/− −/− 24 +/+ +/+ −/− +/+ 48 +/+ +/+ +/+ −/− ↵a RNA was isolated from kidneys of animals inoculated with the specified strain, and RT-PCR was performed using GFP-specific primers. Each entry in the table shows results from a pair of mice at each time point. +, presence of RT-PCR product. −, lack of detectable RT-PCR product. Strains not listed produced all negative reactions. PALS2-GFP and PALS4-GFP reporter strains were not included in the analysis.
- TABLE 3.
Percent positive ALS-specific RT-PCR from analysis of total kidney RNA from mice inoculated with C. albicansa
Duration of infection (h) % Of positive ALS-specific RT-PCR reactions ALS1 ALS2 ALS3 ALS4 ALS5 ALS6 ALS7 ALS9 12 14 0 0 0 0 0 0 0 24 71 86 93 7 0 0 0 7 48 86 93 86 0 0 0 0 14 ↵a RT-PCR was performed with ALS-specific primers (7) on total kidney RNA from the experimental mice. Each data point represents analysis of 14 mice (2 mice each inoculated with CAI12 or one of the PALS-GFP reporter strains). Percentages represent the frequency with which individual ALS-specific transcripts were detected.
