Fig. 4.

MeV-infected cells shed into the apical compartment were detected by microscopy. TEC cultures were infected with WT MeV or LAMV for 4 h and washed with PBS, and media was added. Apical supernatant fluids were collected every 24 h after infection. (A) Cells shed to the apical supernatant fluid after apical infection with Bil-MeV, and LAMV were examined with phase-contrast microscopy or stained with antibody against MeV N after fixation and permeabilization. (Scale bar: 150 µm.) Microscopic examination of shed MGCs after basolateral infection is shown in SI Appendix, Fig. S2. MGCs were seen only after MeV infection but not found in mock-infected cultures. (B) Changes in numbers of shed cells found in rmTEC cultures from 48 to 144 h after apical infection with WT MeV (Bil) and LAMV (EZ). Shed cells were placed on slides by cytospin. Numbers of cells were quantified by counting DAPI-positive nuclei using a confocal microscope. Two-way ANOVA with Tukey’s multiple comparison test comparing mock to infected at each time. **P < 0.05, ***P < 0.001.