Figure 3

Filipin does not affect IPNV infection of CHSE-214 cells. CHSE-214 cells were infected with IPNV at an MOI of 1 for 1 h with 1 or 5 μg/ml of filipin (Fil). After 12 h of incubation at 20°, cells were processed by indirect immunofluorescence (IFI) for IPNV VP2/VP3. Images were recorded with an Olympus Spinning Disk IX81 microscope. For each condition, 250 cells were counted. Number of VP2/VP3-expressing cells is shown as a percentage, normalized to mock cells (A). Control HeLa cells were pre-incubated for 1 h with 0 (B) or 5 μg/ml (C) filipin and 2.5 μg/ml Alexa Fluor 594-conjugated cholera toxin B (CTB-A594) subunit were added. After 30 min, cells were processed for confocal fluorescence microscopy. CHSE-214 cells were preincubated 1 h in the absence (D,E) or with 5 μg/ml filipin (F). CTB-A594 (2.5 μg/ml) was added and cells were incubated for 1 h (D) or 24 h (E and F). Images were recorded with a C2 Plus Eclipse TI Nikon confocal microscope (scale bar = 10 µm). Representative images are shown.