Figure 1

Progression of enteroviral infection in 1.1B4 cells exposed to Coxsackie B4.E2. (A) Cell culture medium collected from 1.1B4 cells was utilised in virus infectivity assays with HEp-2 cells. Viral plaques were examined after plating of culture medium (diluted as shown) that had been collected from mock infected cells (left panel) or from cells on either day 9 (middle plate) or day 29 (right plate) post-infection and incubated on lawns of HEp-2 cells. Representative results from an individual experiment are presented. (B) One Step qRT-PCR was performed with CBV4-specific primers using RNA harvested from culture medium of CVB4.E2 or mock infected 1.1B4 cells. Representative results from an individual experimental run are shown and are presented as relative Ct values (middle panel) in comparison with quantified plaque forming units of virus (left panel). The percentage of dsRNA positive (virally-infected) cells at each time point is shown in the right panel. (C) A representative example of CBV4.E2 or mock infected cells stained using an antibody raised against dsRNA and assessed by flow cytometry at day 64 post infection. The right panel illustrates cells immunostained to detect VP1 and dsRNA (imaged on day 12 post infection).