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. 2021 Sep 27;95(20):e00666-21. doi: 10.1128/JVI.00666-21

FIG 7.

FIG 7

Linker-HA tags abrogate fusogenicity of headless NiV G and HeV G. (A) Schematic representation of HA-tagged NiV G/HeV G and linker-HA tagged NG167(HA)/HG167(HA). The protein linker is a (GGGGS)3 motif designed to retain the function of both linked proteins, headless G and HA tag (30). Note that all wild-type G used throughout the study had a C-terminal HA tag even if not shown in other schematics. (B) Relative levels of fusion for wild-type or headless linker-tagged NiV G and HeV G coexpressed with cognate F in HEK 293T cells at 16 to 20 hpt normalized to NG/NF set to 100% (mean ± SD, n = 3). (C) Relative levels of G CSE for NG167(HA)/NF and HG167(HA)/HF normalized to NG167(HA)/NF set to 100% (mean ± SD, n = 3). (D) Representative blots of coimmunoprecipitated supernatants and lysates of NG167(HA)/NF and HG167 (HA)/HF (24 hpt). The leftmost lanes represent the co-IP negative control. (E) Representative fusion images (outlined) for NG167(HA)/NF and HG167(HA)/HF in HEK 293T cells (24 hpt). All signals were obtained from the same blot, but the lanes were not necessarily adjacent. Gray points in the plots represent individual data points.