B cell receptor revision diminishes the autoreactive B cell response after antigen activation in mice
J. Clin. Invest. Ying-Hua Wang, et al. 118:2896 doi:10.1172/JCI35618 [Go to this article.]

Figure 1
RAG is induced in antigen-reactive early memory/preplasma B cells. (A) Histological staining of spleen sections for peanut agglutinin (PNA) (red) and DWEYS-tetramer (green, left 2 panels) or RAG2 (green, right panel). Original magnification: ×100 (left and right panels); ×400 (middle panel). (B) Histological staining of spleen sections for B220 (red), tetramer binding (green), and RAG2 (blue). RAG2 is coexpressed in extrafollicular tetramer-binding cells. Original magnification: ×100 (top row); ×400 (middle and bottom rows). Three to five mice were used in each group. Four to six pictures were taken for each spleen section. (C) Bivariate plots showing the gates for sorting the specified B cell subsets. (D) qPCR analysis of RAG1 and RAG2 expression in antigen-reactive and nonreactive B cells. BM immature B cells (B220⁺AA4.1^hiIgM⁺IgD^–) were used as a positive control. Mice were immunized with DWEYS-MAP in CFA on day 0 and boosted with DWEYS-MAP in incomplete Freund adjuvant on day 7. On day 16, specified B cell subsets were sorted by flow cytometry for preparation of RNA. RNA polymerase 2a (Polr2a) was used as internal control gene. Data (mean ± SEM) are representative of at least 4 independent experiments. Relative level is defined as the relative expression of mRNA normalized to that of Polr2a.