Fig. 5
Stimulation of avian macrophages with the synthetic ssRNA, resiquimod increases the production of IL-1β but not type 1 IFNs. a Avian macrophages, MQ-NCSU cells, were cultured in 12-well plates (1 × 106 cells per well) for 24 h and stimulated with either resiquimod (10 μg/ml) or growth media (control) in triplicates. The culture supernatants were collected 24 h post-treatment and transferred to DF-1 cell monolayers grown in 12-well plates and infected with VSV conjugated with GFP (0.1 MOI). At 24 h post infection, the percentage of cells expressing GFP signal was quantified after fixation with 4% paraformaldehyde and nuclear staining. The experiment was repeated two more times with similar results and the data were pooled. The representative figures in each group are shown. b The MQ-NCSU cells were cultured on coverslips in 12-well plates with 1 × 106 viable cells per well and protein transport inhibitor (2 μl/ml) was added to culture medium after 6 h. After 24 h of culture, the cells were stimulated with resiquimod (10 μg/ml), or RPMI media as a control separately (triplicates per treatment). Following 24 h of treatment, immunofluorescent staining targeting IL-1β was performed after fixation with 4% paraformaldehyde. The experiment was repeated two more times with similar results and the data were pooled. The representative figures in each group are shown. Student’s t-test was used to identify differences between two groups and the differences were considered significant at P < 0.05