Growth curve and multicycle Influenza infection measurements

For multicycle influenza A infection experiment, confluent A549 cells were infected in a 12 well plate format with A/Victoria/3/75 virus at a MOI of 0.001 PFU/cell and then incubated for 1h at 37°C and 5% CO₂. The inoculum was removed, cells were washed once with 1ml PBS and 1ml of serum-free DMEM with 0.2ug/ml of TPCK-treated trypsin containing either Mito-C (2μM), Oseltamivir (50nM) or DMSO was added before incubating cells at 37°C. Infections were performed in triplicates. At 24, 48, 72 and 96h post-infection, samples were collected and frozen for viral titration by standard plaque assay on MDCK cells.