Journal: Proceedings of the National Academy of Sciences of the United States of America
Article Title: BRCT-domain protein BRIT1 influences class switch recombination
Figure Lengend Snippet: BRIT1 is dispensable for immune response in vivo. ( A ) Schematic of NP-CGG immunization protocol. GC, germinal center. ( B ) IgM, IgG1, IgG3, and IgA concentrations in serum, determined by ELISA on day 0. Ctrl, control. ( C ) Flow cytometric analysis of GC B cells (B220 + DAPI − GL7 + Fas + ) in control, BRIT1 KO, and AID KO mice following NP-CGG immunization on day 14 and quantification of GC B cells in immunized mice on day 14 ( n = 3). ( D ) Quantification of IgG1 + GC B cells in immunized mice on day 14 ( n = 3). ( E ) Quantification of NP + GC B cells in immunized mice on day 14 ( n = 3). ( F ) Quantification of IgG1 + NP + GC B cells in immunized mice on day 14 ( n = 3). ( G ) IgM and IgG1 concentrations in serum from Ctrl, BRIT1 KO, and AID KO mice following NP-CGG immunization on day 28, determined by ELISA. ( H ) Detection of low-affinity and high-affinity NP-specific IgM and IgG1 serum antibodies in Ctrl ( n = 4), BRIT1 KO ( n = 4), and AID KO ( n = 4) mice following NP-CGG immunization on day 28 by ELISA. Conc., concentration.
Article Snippet: The following isotype standards were used to calculate absolute concentration values: IgM (no. 14-4752-81; eBioscience), IgG1 (no. 0102-01; Southern Biotechnology Associates), IgG3 (no. 553486; BD Pharmingen), and IgA (no. 553478; BD Pharmingen).
Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Mouse Assay, Concentration Assay