The WT protein and NxT PNGS mutants were analyzed by negative stain EM as previously described (Cottrell et al., 2020 (link)). For the RM20E1 complexes, SOSIP trimers were incubated with 6-fold molar excess per protomer Fab at RT overnight. The complexes were purified using SEC on a Superose 6 Increase 10/300 GL (GE Healthcare) column. Samples were imaged and analyzed as previously described (Cottrell et al., 2020 (link)).
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