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. 2023 Feb 27;11(2):e03637-22. doi: 10.1128/spectrum.03637-22

FIG 6.

FIG 6

Silencing circMerTK enhances the innate antiviral response, resulting in impaired expression of the IAV NP gene. (A) NIH/3T3 cell lines stably expressing specific shRNAs targeting circMerTK or luciferase were infected with or without IAV PR8 for 14 h. IFN-β, MX1, and IFITM3 mRNA expression levels were determined by RT-PCR. Western blotting was utilized to assess IAV-NP levels. (B) mRNA levels of IFN-β, (C) MX1, and (D) IFITM3 were evaluated in circMerTK knockdown NIH/3T3 cell lines by RT-qPCR as described in panel A. (E) A549 cell lines stably expressing specific shRNAs targeting circMerTK or luciferase were infected with or without IAV PR8 for 14 h. IFN-β, MX1, and IFITM3 mRNA expression levels were determined by RT-PCR. Western blotting was utilized to assess IAV-NP levels. (F) circMerTK knockdown A549 cells, along with the control, were infected with IAV PR8 for 16 and 18 h. Enzyme-linked immunosorbent assay (ELISA) was used to examine IFN-β protein levels in the supernatants at the indicated time points. (G) mRNA levels of ISG15 and (H) IFITM3 were evaluated in circMerTK knockdown A549 cell lines by RT-qPCR as described in panel E. Representative data from three independent experiments are shown. Data represent the mean values ± SD (n = 3; *, P < 0.05; **, P < 0.01).