Fig. 4

Motility of podocytes incubated with supernatants of infected podocytes. a Untreated, UV-irradiated, and filtrated supernatants of podocytes infected with PUUV for six days at an MOI of 0.5 were analyzed for the presence of N protein by Western blot and for infectivity via incubation of Vero E6 cells and subsequent detection of N protein (red). Cells were imaged at a magnification of × 200. b Supernatants and filtrates were added to uninfected podocytes 30 min before migration assay. Migration assay was performed in the presence of supernatants and filtrates and analyzed as described in Fig. 2c. Migration of podocytes in the presence of untreated, UV-irradiated, and filtrated supernatants of uninfected podocytes served as control (ctrl) and was set to 100%. (c and d) Supernatants of HTNV-infected podocytes were treated and analyzed as described for PUUV in (a and (b). (e and f) Time course of infection and motility analyzed in podocytes inoculated with untreated supernatants derived from HTNV-infected podocytes after six days of infection. e Infection was quantified by counting cells expressing N protein at the indicated time points. f Migration assays were started after 30 min of incubation with supernatant and motility was analyzed at the indicated time points. Untreated supernatants of uninfected podocytes served as control (ctrl) and was set to 100%. Results were obtained from three independent experiments. Shown is mean ± SD