Fig. 3
From: Cellular investigations with human antibodies associated with the anti-IgLON5 syndrome
Deglycosylated IgLON5 is recognized by human IgLON5 antibodies. A Anti-IgLON5 immunoreactivity in rat brain sections is totally abrogated when the anti-IgLON5-positive serum is preabsorbed with glycosylated (a) or deglycosylated protein extracts from transfected HEK293 with IgLON5 (c). The immunoreactivity of IgLON5 antibodies is present when the serum is preabsorbed with an unrelated antigen glycosylated (b) or deglycosylated (d). Scale bar = 1000 μm. B a Immunofluorescence of HEK293 cells transfected with IgLON5. Prevention of N-glycosylation with tunicamycin treatment does not affect the reactivity of patients’ antibodies with IgLON5. Scale bar = 10 μm. b Western blot of extracts of the indicated IgLON5-HEK293 cells shown in (a), demonstrating a shift of the molecular weight (from ~52 to 36 kDa) consistent with the deglycosylation of the protein. The findings confirm that the reactivity of patients’ serum antibodies with tunicamycin-treated cells of A was directed against deglycosylated epitopes