Figure 1.

A. fumigatus Conidia Induce IFN-γ-Dependent DAPK1 Expression In Vitro and In Vivo

(A–C) DAPK1 gene expression (A) and production by immunoblotting (B) and immunofluorescence (C) in RAW264.7 cells pulsed with live A. fumigatus conidia in the presence of rIFN-γ.

(D) DAPK1 expression in C57BL/6 lung macrophages after exposure to A. fumigatus heat-inactivated resting conidia (RC), swollen conidia (SC), or hyphae.

(E and F) DAPK1 gene (E) and protein (F) expression in lung of mice either uninfected (0 dpi, days post-infection) or infected with A. fumigatus conidia and treated with rIFN-γ.

(G) Immunoblotting of DAPK1 in purified lung macrophages exposed to live A. fumigatus conidia in vitro in the presence of rIFN-γ. Dapk1 gene expression was assessed by real-time qPCR and normalized on Gapdh.

For immunoblotting, normalization was performed on mouse β-actin (the corresponding pixel densities are indicated). For immunofluorescence, nuclei were counterstained with DAPI. Photographs were taken with a high-resolution microscope (Olympus BX51). Scale bars, 25 μm. DAPK1 fluorescence intensity was measured with the ImageJ software; BF, brightfield. Data (mean values ± SD) represent pooled results or representative images (immunofluorescence and immunoblotting) from three experiments. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001, rIFN-γ treated versus untreated RAW264.7 cells and rIFN-γ treated versus untreated Ifng^−/− mice. None, unpulsed and/or untreated cells. See also Figure S1.