Mice were infected with EMCV by intraperitoneal injection of 2×103 TCID50 of the virus diluted in 100 μl PBS. Mice were sacrificed and heart harvested 48h later, or survival was monitored for 27 days.
For VSV infection, mice were anesthetized with methoxyflurane (Anafane) and 106 PFU of VSV in 50 μl PBS were administered intranasally. The mice were sacrificed 24h later. Blood was collected by cardiac puncture and transferred in heparinized tubes. The samples were then centrifuged (2 minutes at 3000 rpm) and dilutions of the plasma were used for viral titration.
For in vitro infections, cells were plated at a density of 105 cells/ml in 6-well plates. The next day, the cells were infected with VSV-GFP, VSV-DsRed, HSV-2 or EMCV (diluted in 500 μl of DMEM without FBS) for 1h with gentle shaking every 15 minutes. The cells were then washed and incubated in 1 ml of media containing 10% FBS. The dose of virus used for infection and the duration of the incubation are indicated in figure legends. Cell death was measured by flow cytometry after staining with Annexin V-APC and propidium iodide (BD Biosciences) or by LDH release assay (CytoTox 96 assay, Promega).
For VSV infection, mice were anesthetized with methoxyflurane (Anafane) and 106 PFU of VSV in 50 μl PBS were administered intranasally. The mice were sacrificed 24h later. Blood was collected by cardiac puncture and transferred in heparinized tubes. The samples were then centrifuged (2 minutes at 3000 rpm) and dilutions of the plasma were used for viral titration.
For in vitro infections, cells were plated at a density of 105 cells/ml in 6-well plates. The next day, the cells were infected with VSV-GFP, VSV-DsRed, HSV-2 or EMCV (diluted in 500 μl of DMEM without FBS) for 1h with gentle shaking every 15 minutes. The cells were then washed and incubated in 1 ml of media containing 10% FBS. The dose of virus used for infection and the duration of the incubation are indicated in figure legends. Cell death was measured by flow cytometry after staining with Annexin V-APC and propidium iodide (BD Biosciences) or by LDH release assay (CytoTox 96 assay, Promega).