Figure 5

Polar entry and release of HTNV from MDCKII cells. A) Polarized MDCKII monolayers were infected with HTNV and analyzed for the expression of nucleocapsid protein by immunofluorescence. B) MDCKII cells were inoculated with HTNV via the apical or basolateral chamber. MDCKII cells were lysed and hantaviral nucleocapsid protein was detected by Western blot. Equal loading was monitored by detection of tubulin. C) MDCKII cells were inoculated with HTNV via the apical or basolateral chamber. Cell-free supernatants of the apical and basolateral chamber of infected polarized MDCKII monolayers were analyzed for the presence of nucleocapsid protein by Western blot. D) Apical and basolateral cell-free culture media of infected MDCKII cells were added to Vero E6 cells and re-infection was monitored by the detection of viral nucleocapsid protein.