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Figure 2 | Journal of Neuroinflammation

Figure 2

From: A novel autoantibody against ephrin type B receptor 2 in acute necrotizing encephalopathy

Figure 2

Identification of ephrin type B receptor 2 (EphB2) as a novel autoantigen. (A) Nonpermeabilized human umbilical vein endothelial cells (HUVECs) were stained with 0.5 mg/mL of IgG of control or V10-5 followed by secondary antibody and analyzed by flow cytometry. (B) HUVEC cDNA-expressing cells were stained with 0.5 mg/mL of V10-5 IgG followed by secondary antibody, and cells in the positive fraction were sorted (squares). (C) Unsorted cells and those at the fourth round of sorting (upper), and unsorted and cloned cells from those at the fourth round of sorting, C17 (lower), were stained with 0.5 mg/mL of V10-5 IgG followed by secondary antibody, and analyzed by flow cytometry. (D) EphB2 cDNA fragments inserted into the genomic DNA of C17 were amplified, and PCR products were electrophoresed on a 0.8 % agarose gel. (E) Unsorted and C17 were stained with isotype control or anti-EphB2 antibody, followed by secondary antibody and analyzed by flow cytometry. (F) Expression vector, empty-IRES-GFP, or EphB2-IRES-GFP was transfected into YB 2/0 cells, and the cells were stained with 0.5 mg/mL of control IgG or V10-5 IgG, followed by secondary antibody and analyzed by flow cytometry.

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