Fig 1: Chitin accumulation on the surface of F. pedrosoi-sclerotic cells compensatorily increased after chitinase treatment in a time-dependent manner.(A-F) Abbreviations: F. pedrosoi-spore (F. p-spore); F. pedrosoi-hyphae (F. p-hyphae); In-vitro transformed F. pedrosoi-sclerotic cells in ATCC 830 medium with 10−6 M PAF for 50 days (FPSC); Heat-killed FPSC (HK-FPSC); Chitinase (CHIA). (A and B) The binding capacity of murine Dectin-1 to F. p-spores, F. p-hyphae, FPSC or HK-FPSC was respectively detected by PE-conjugated anti-murine Dectin-1 mAb using flow cytometry, and was represented as mean fluorescence intensity (MFI). (C and D) The binding capacity of murine Dectin-2 to F. p-spores, F. p-hyphae, FPSC or HK-FPSC was respectively detected by PE-conjugated anti-murine Dectin-2 mAb using flow cytometry, and was represented as mean fluorescence intensity (MFI). (A-D) The fungal cells incubated only with PE-conjugated isotypes were set as blank control. (E and F) The binding capacity of FITC-conjugated WGA to F. p-spore, F. p-hyphae, FPSC or HK-FPSC before and after chitinase treatment at indicated time points was detected by flow cytometry, and was represented as mean fluorescence intensity (MFI). (B, D and F) Data represent the mean±SEM from three individual experiments performed in triplicates, and statistical analysis was performed using Univariate ANOVA and LSD-t test. Significant: * P<0.05; Highly Significant: ** P<0.01.
Fig 2: Dectin-1, MR and Dectin-2 mainly bind to the surface of F. pedrosoi-hyphae, but not the transformed sclerotic cells.(A-C) The binding of murine-derived Dectin-1, Mannose Receptor (MR), and Dectin-2 to F. pedrosoi cultured in ATCC 830 medium with 10−6 M PAF at 35°C for 30 days was detected respectively by PE-conjugated anti-murine Dectin-1 mAb (A), FITC-conjugated anti-murine MR mAb (B), and PE-conjugated anti-murine Dectin-2 mAb (C) using confocal microscope. Left column: fluorescence field; Middle column: bright field; Right column: merged images. Scale bar = 20 μm. The transformed sclerotic cells with cross-septation and swelling chlamydospores were indicated by red circle (A) or red arrows (B and C). The fungal cells incubated only with PE-conjugated anti-murine Dectin-1 or Dectin-2 mAb, or only with FITC-conjugated anti-murine MR mAb were set as blank control.
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