Fig 2

TRAF3 overexpression promotes lung epithelial cell adhesion and internalization of A. fumigatus spores. A549 cells were transfected with pIRES-TRAF3-EGFP plasmid (TRAF3) and control pIRES-EGFP plasmid (Vector). mRNA (A) and protein (B, C) expression of TRAF3 were determined by qRT-PCR and western blot. A. fumigatus spores were co-incubated with A549 cells with or without TRAF3 overexpression for 6 h. A. fumigatus spores were labeled with fluorescent white (CFW), then fixed with 4% paraformaldehyde. Cells were dead after fixation and then Annexin-V-stained cell membrane; the number of A. fumigatus spores adhered to lung epithelial cells was determined by fluorescence (D). (E) is the number of A. fumigatus spores adhered to each cell. The number of internalized A. fumigatus spores with or without TRAF3 overexpression in A549 cells was determined by nystatin protection assay (F). Results are expressed as the mean ± SD of three independent experiments. **P < 0.01, ***P < 0.001. Scale bars: 50 µm.