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. 2021 Mar 23;101(3):971. doi: 10.2340/00015555-3774

Fig. 2.

Fig. 2

Schematic representation of type VII collagen (Col7) and the Col7 domains used in the different serological tests as well as non-collagenous NC1 BIOCHIP® and immunoblotting (IB). (a) The NC1 and/or NC2 domain of Col7 or full-length Col7 are used in the different serological assays for the detection of Col7-specific immunoglobulin G (IgG) autoantibodies. (b) Exemplified pictures of an indirect immunofluorescence (IF) microscopy using the NC1 BIOCHIP®. The recombinant NC1 domain is expressed on the surface of HEK293 cells (red box, left field). HEK293 transfected with the empty plasmid (pTriEx-1) served as control (red box, right field). A representative picture of a positive epidermolysis bullosa acquisita (EBA) serum reacting with NC1-expressing HEK293 cells is shown (red box, left field). (c) Representative pictures of immunoblotting using extracts of human dermis (upper blot) and recombinant NC1 (lower blot). Upper blot: lane 1, anti-p200 pemphigoid serum labelling the p200 protein; lane 2, EBA serum labelling the 290 kilodalton (kD) full-length Col7 protein; lanes 3–4, sera of healthy blood donors; lower blot: lanes 1–2, sera of patients with EBA reacting with the NC1 domainofCol7, lanes3–5,seraof healthy blood donors. H2N: N-terminus; COOH: C-terminus; CMP, cartilage matrix protein (CMP) domain; vWFA2, von Willebrand factor A2-like domain; 1-9, nine fibronectin III (FNIII)-like domains; BPTI: Kunitz bovine pancreatic trypsin inhibitor domain.