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. 2021 Jul 22;218(9):e20210580. doi: 10.1084/jem.20210580

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© 2021 Cavazzoni et al.

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Figure S5. — Antigen specificity of B cells in GCs after immunization with OVA. BALB/c mice were immunized s.c. with OVA (2 μg/mouse) combined with R848 (1 μg/mouse). When indicated, a booster immunization was performed on day 7 after prime. (A) Kinetics of serum levels of IgG binding to OVA, measured by ELISA. O.D. sum is the summation of ODs of four serum dilutions (1:40, 1:120, 1:360, and 1:1,080). (B and C) Single GC B cell (B) or pooled GC B cells (C) were sorted and cultured on a monolayer of gamma-irradiated NB21 feeder cells (10³ cells/well; Kuraoka et al., 2016). After 7 d, supernatants were collected for binding assays. Supernatants were screened for IgG production and IgG⁺ wells were tested for binding to OVA protein by ELISA or immunoblot against mouse brain tissue as source of self-antigens. Ctrl, serum from ZIKV infected mice. One experiment was performed with three mice.