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. 2020 Mar 9;11:1253. doi: 10.1038/s41467-020-15088-0

Fig. 1. Subcloning of autoantigen-expressing cells by using IgG from patients with Takayasu arteritis.

Fig. 1

a AECA activity against HAECs or HUVECs treated with or without 1 ng/mL TNF-α for 6 h was measured with flow cytometry in patients with Takayasu arteritis (TAK, n = 21). Dots represent the data for individual subjects. Dots connected with lines indicate the same patients. Control indicates healthy individuals (n = 79). b Nonpermeabilized HUVECs were stained with 0.5 mg/mL purified IgG obtained from a healthy individual or patients with TAK, followed by incubation with secondary antibody and flow cytometry analysis. c–h Sorting of cells expressing cell-surface autoantigens by using three different AECAs. c, e, g YB2/0 cells expressing HUVEC cDNA were stained with 0.5 mg/mL U10-4 IgG (c), W10-59 IgG (e), or G10-43 IgG (g), followed by incubation with secondary antibody, and cells in the positive fraction (squares) were sorted by flow cytometry. FITC-conjugated IgG antibody was initially used, and PE-conjugated IgG antibody was subsequently used as a secondary antibody. d, f, h Binding activity of serum IgG to unsorted cells or cloned cells from sorted cells (C1 [upper] and C3 [lower] isolated from U10-4, D; C6 isolated from W10-59, F; C7 isolated from G10-43, H, respectively); cells were stained with 0.5 mg/mL prototype AECA IgG, followed by incubation with secondary antibody and flow cytometry analysis. G10-43, W10-59, and U10-4 indicate the serum sample number.