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. Author manuscript; available in PMC: 2019 Jul 1.
Published in final edited form as: Nat Microbiol. 2019 Feb 18;4(5):837–845. doi: 10.1038/s41564-019-0369-x

Figure 5: Inter- and intra-layer interactions in NS1 tubules.

Figure 5:

(a) Interlayer six-monomer junction in NS1 tubule. Blue inset, atomic models showing intra-layer beta-sheet augmentation involving the head domain of one monomer and the C-terminal arm of the other monomer. Yellow inset, atomic models of sample intra-layer packing between the head domain of one monomer and the foot domain of another monomer. Monomer-monomer interfaces A and B are indicated. (b) Putative disulfide bond between monomers of the same layer. Black inset: atomic models of C364 sidechain position and orientation. (c) Recovery of C364S mutant virus by reverse genetics as compared to the wild-type virus. Plaques (white) indicative of recovered viruses, were visualized by crystal violet staining. Similar plaque phenotypes were observed for the wild-type and mutant viruses. (d) Intracellular localization of wild-type or C364S mutant NS1. Immunofluorescence analysis showed similar VIBs (indicative of similar viral recovery) but partially affected intracellular NS1 localization in cells infected with the mutant virus as compared to those infected with the wild-type virus. NS1, green (rabbit anti-NS1 primary antibody and anti-rabbit Alexa 488 coupled secondary antibody); NS2, red [component of the viral inclusion bodies (VIBs), mouse anti-NS2 primary antibody and anti-mouse Alexa 546 coupled secondary antibody]; Nuclei, blue (Hoechst staining). Scale bars, 20 μm. (c, d) Experiments were repeated twice independently with similar results.