Figure 7
BHV-1-GFP and BRSV-GFP infection of PCLS. One day after preparation, PCLS were inoculated with BHV-1-GFP for 2 h or BRSV-GFP for 3 h (105 FFU/mL). BHV-1-GFP infection (A-C) was stopped after 2 days (A, B) or after one week (C) and cells were stained for β-tubulin (A, C) to detect cilia or with anti-CD-142 for detection of basal cells (B, both in red). BRSV-GFP infection (D-F) was stopped after 3 days (D, E) or one week (F) and slices were stained for β-tubulin (red) and DAPI (blue). (D) BRSV-infected cells detected in the peribronchiolar connective tissue; (E) a characteristic shape of a BRSV-GFP-infected cell in the submucosal region. F) BRSV-GFP-infected cells within the peripheral part of the slice. The insert shows infected cells within the epithelium lining the alveolar lumen. GFP-expressing cells indicate virus-infected cells. Scale bars = 25 μm in (A + E) and 100 μm in (B-D, F).