Extended Data Fig. 7: Fucosylaton of GP2.
From: Bacterial and host fucosylation maintain IgA homeostasis to limit intestinal inflammation in mice
Fucosylaton of GP2 mediates the interaction of IgA in in the follicle-associated epithelium (FAE) of PPs. (a) Co-immunoprecipitation (co-IP) of IgA and GP2. Immunocomplexes were co-IP from small intestine IEC lysate of WT mice or Fut2-/- mice with anti-GP2 antibody and then were stained for immunoblotting of IgA by anti-IgA antibody. Insert digital: Mean Density. (b) Cross-section immunostaining of GP2 (green) and UEA-1 (red) in the follicle-associated epithelium (FAE) of PPs. Scale bars, 50 μm. (c) Cross-section immunostaining of GP2 (green), IgA (purple) and UEA-1 (red) in the follicle-associated epithelium (FAE) of PPs of WT mice or Fut2-/- mice. Scale bars, 50 μm. (d) Co-IP of IgA and GP2. IgA-/- ileal lysate was incubated with biotinylated anti-mouse GP2 antibody and captured using streptavidin agarose beads. For fucosidase O treatment groups, beads capturing GP2 were incubated with 10 U of fucosidase O at 37°C for 6 hours prior to IgA pulldown. The beads were washed and subsequently incubated with WT ileal lysate to allow IgA binding. Bound proteins were eluted and analyzed by Western blotting using anti-IgA-HRP antibody. The data are representative of two independent experiments (a-d).
