AmiRs inhibit CVB3 replication in AmiR-expressing cell lines.

<p><b>A) Western blotting detection of CVB3 VP-1 protein.</b> Different AmiR cell lines were infected with CVB3 (Kandolf) and cell lysates were collected for VP-1 detection by Western blotting. Sham infected cells served as a negative control. The VP-1 expression levels were quantified by densitometry and normalized to the level of beta-actin. The ratios of VP-1 to beta-actin were further converted to the ratios of the value of AmiR-S cells, which was set as 1.0. <b>B) Viral plaque assay.</b> Cells were infected with CVB3 at 0.01 MOI for 24 h. The supernatant was collected to perform viral plaque assay to determine the viral titer. The upper panel is the representative viral plaques for each cell line and the lower panel is the calculation of viral titers. <b>C) q-RT-PCR detection of mature AmiRs.</b> Total RNAs from the AmiR-1, AmiR-2 and AmiR-S cell lines were extracted. The expression of mature AmiR-1 and AmiR-2 were measured by q-RT-PCR and normalized to the expression of U6 RNA in each sample. The relative mature AmiR-1 and AmiR-2 levels in the AmiR-S cell line were set as 0. The values shown represent the means ± SD of the data from three independent experiments, <i>P</i><0.05.