Figure 4.

Analysis of the N-glycosylated GB2 mutants. (A) Western blot analysis of the full-length Flag–GB2 mutants from membrane fractions of cells coexpressing Flag–GB2 mutants and HA–GB1_ASA. As indicated in Figure 3, cartoons depict the GABA_B subunits wild-type (white) and mutant (grey), and engineered N-glycan is indicated by a white star. (B) Western blot analysis of the truncated GB2 subunits deleted of both HD and C-terminal regions, with or without treatment with PNGase F, and comparison to the wild-type construct. (C) The amount of Flag–GB2-WT mutants coexpressed with HA–GB1_ASA at the cell surface as measured by ELISA. (D) IP production for the Flag-tagged GB2 mutants coexpressed with GB1_ASA. Data are means±s.e. of at least three independent determinations.