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. 2019 Apr 29;129(6):2222–2236. doi: 10.1172/JCI124694

Figure 1. Antibodies to Nfasc155 do not alter the interaction between Nfasc155 and its axonal partners CNTN1 and CASPR1.

Figure 1

(A–C) For aggregation assays, HEK293 cells were cotransfected with mCherry-tagged Nfasc155 or with GFP-tagged CASPR1 and CNTN1. Cells were then incubated under gentle agitation for 2 hours in the presence of 10 μg of control IgG4, anti-CNTN1 IgG4, or anti-Nfasc155 IgG4 from 3 patients (CIDP1–3). As negative controls, Nfasc155-expressing HEK293 cells were incubated with cells expressing GFP alone (top left panel). Cells were examined with a fluorescence microscope at ×10 objective. Representative fields are shown in A (n = 3–4 experiments for each condition). Dashed circles highlight cell aggregates with contacts between red and green cells. The percentage of cell clusters with contacts between green and red cells was quantified (B), as well as the relative frequency of green cells per aggregate (C) (n = 3–4 experiments for each condition). CASPR1/CNTN1– and Nfasc155-expressing cells form clusters. Anti-CNTN1 IgG4 significantly prevented the formation of cell aggregates (**P < 0.005 by unpaired 2-tailed Student’s t tests for 2 samples of equal variance and by 1-way ANOVA followed by Bonferroni’s post hoc tests). By contrast, anti-Nfasc155 IgG4 did not affect the interaction between Nfasc155 and CASPR1/CNTN1. Bars represent mean and SEM. Scale bar: 50 μm.