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. 2019 Aug 16;6(5):e603. doi: 10.1212/NXI.0000000000000603

Figure 3. Epitope mapping of anti-NF autoantibodies by binding assays to truncated NF mutants and flow cytometry.

Figure 3

The NF isoforms consist of a common Ig6 domain and different isoform specific fibronectin domains (A, right section). Reactivity of patient sera to NF-155 and NF-186 (A) and truncations of NF (B) by flow cytometry: patients 1, 2, and 3 show reactivity to both NF-155 (blue line) and NF-186 (orange line) compared with control (empty RSV vector, black, filled), whereas patients 4 and 5 only react to NF-155 (A). Epitope analysis using NF truncations (B) demonstrated that reactivity was against the 6Ig domain in 3 of the patients (patients 1–3) who were positive for both NF isoforms (green line). Patients 1 and 2 also reacted to the Fn3 domain (red line). The Fn3Fn4 domain (yellow line) was recognized by the sera of patients 4 and 5, who reacted only to NF-155. Other truncated NF variants (Fn1Fn2 = dark blue; Fn4-Mucin-Fn5 = brown; Mucin-Fn5 = purple; GFP-only control = gray filled) were not recognized by any of the sera tested (sera 1–4 and 6 tested for all mutants, serum 5 for Fn3Fn4 only). NF = neurofascin.