Fig. 2

Phillyrin suppressed influenza viral replication and reduced influenza virus-induced cytopathic effect (CPE). Madin Darby Canine Kidney (MDCK) cells were inoculated with IAV (PR8) at multiplicity of infection (MOI) 0.005. After 2 h adsorption, virus inoculum was removed and replaced with freshly prepared infectious media with phillyrin at different concentrations (100, 50, 25µM) or ribavirin at 10µM (as a positive control). Viral RNA (M mRNA and NP mRNA ) replication (a-d): Cells were collected at12hpi and 24hpi respectively, and RNA was isolated, and M mRNA and NP mRNA were determined using quantitative real-time polymerase chain reaction (qRT-PCR). % of M mRNA levels (a and c) and NP mRNA levels (b and d) in virus-infected samples in the presence of Phillyrin relative to virus infection alone was calculated. The graphs represent the mean and standard deviation of three independent experiments of treatment by Phillyrin. Significant differences were identified by one-way analysis of variance (ANOVA), *p < 0.05, relative to virus infection alone without treated group. Virus-induced CPE was recorded under microscopy at 48hpi (e-j). Results shown are from a typical experiment.“Normal Control”is the negative control with no influenza virus, cultured for the same time period in the presence of phillyrin and ribavirin