Fig. 5
dsRNA treatment of macrophages elicits antiviral response against LPAIV infection and type I IFN activity. Avian macrophages were cultured in 6- well plates or 12- well plates at 1.5 × 106 or 7.5 × 105 cells per well respectively for 24 h and then, treated with dsRNA at 50 μg/ml (n = 3) or RPMI medium as control (n = 3) for 24 h. (a) MQ-NCSU cells infected with H4N6 LPAIV (MOI = 0.1) for 24 h, supernatants were collected and plaque assay was done on MDCK cells. (b) For type 1 IFN activity measurement, the MQ-NCSU cell monolayers were infected with VSV-GFP (MOI = 0.1) for 24 h and the plates were scanned by In Cell Analyzer 2000 machine for green fluorescent signals and further analyzed by In Cell Analyzer 1000 software. Student’s t-test was done to identify group differences and the differences were considered significant at P < 0.05. The data are expressed as the mean ± SEM and the results represents pooled data of 3 independent experiments. The representative images of nuclear stained, VSV-GFP positive and merged pictures are also given for each group