Respiratory Syncytial Virus Nonstructural Protein 1 Promotes 5-Lipoxygenase via miR-19a-3p

Figure 1

RSV NS1 protein upregulates the expression of miR-19a-3p in A549 cells. (a) Western blot analysis of NS1-FLAG protein in A549 cells transfected with pNS1. Anti-FLAG antibodies were used to assess for the presence of FLAG-tagged NS1. β-Actin was used as an internal control. (b) The expression levels of miR-19a-3p mRNA at different time points after pNS1 transfection. U6 RNA was used as an internal control. (c) Localization of NS1 (green) in A549 cells transfected with pNS1 was analyzed by immunofluorescence confocal microscopy at four hours after transfection. Anti-FLAG antibodies were used to assess for the presence of FLAG-tagged NS1. Nuclei were counterstained with DAPI (blue). Representative images were shown. (d) The expression levels of miR-19a-3p mRNA in A549 cells at different time points after RSV inoculation. Monolayers of A549 cells were infected for 72 hours with ultraviolet-inactivated RSV or live RSV. Cells were pretreated with siNC or siNS1 for 6 hours prior to RSV inoculation. U6 RNA was used as an internal control. Data are expressed as ( technical replicates). The experiment was performed three times with similar results, and one representative experiment is shown. vs. 0 h. NS1: nonstructural protein 1; pNS1: nonstructural protein 1 expressing plasmid; DAPI: 4,6-diamidino-2-phenylindole; RSV: respiratory syncytial virus; UV-RSV: ultraviolet-inactivated RSV; siNC: negative control siRNA; siNS1: RSV NS1 siRNA.

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