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. 2019 Dec 23;219(2):e201903062. doi: 10.1083/jcb.201903062

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© 2019 Ci et al.

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Figure 5. — NS1 functions as a dimer. (A) Schematic showing the hetero-dimerization between Flag-tagged WT and Myc-tagged NS1. (B) Dimerization of NS1 proteins with different tags in cotransfected cells. Flag-tagged WT NS1 and Myc-tagged WT or mutated NS1 was coexpressed in HeLa cells. Immunoprecipitation (IP) was performed using a Myc antibody. The proteins precipitated by Myc-tagged NS1 were immunoblotted (IB) with a Flag antibody. (C and D) NS1 mutants interfere with WT NS1-induced ER remodeling. Cells were cotransfected with Myc-tagged WT or mutated NS1 and Flag-tagged WT NS1, and then immunofluorescence was examined. ER morphology is indicated by NS1 staining. Scale bar, 20 µm. (D) The percentage of ER-remodeled cells was defined as the ratio of the number of cells with ER remodeling to the number of cotransfected cells (green and red double-labeled). The data are presented as the mean ± SEM. The P values are obtained from a two-tailed t test. ***, P < 0.001. (E) NS1-mutated replicons interfere with the luciferase activity of WT replicons. BHK-21 cells were cotransfected with WT and NS1-mutated replicons, and the luciferase activity was measured at 10 h and 36 h after transfection.