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. 2007 May 14;204(5):1145–1156. doi: 10.1084/jem.20061952

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Copyright © 2007, The Rockefeller University Press

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Figure 5. — Permanent labeling of B cells expressing AID. (A) Upper schematics: molecular maps of the AID-Cre (top) and Rosa-Neo-EYFP (bottom) transgenes (S, SalI; P, PacI; E, EcoRI). Lower Southern blot: three independent AID-Cre transgenic lines were generated, with copy number ranging between 1 to 5 copies. (B) FACS pseudocolor plots showing CD95-PE and YFP expression in B cells (gated on B220 (PercP-Cy5.5)) from AID-Cre-YFP compound mice. Left panel: axillary lymph node cells from nonimmunized mice; right panel: PPs. (C) Immunofluorescence analysis of gut lamina propria CD138⁺ plasma cells from AID-Cre-YFP mice. Scale bar = 20μm. (D) Mutation analysis of the J_H4 intron from CD95^lowYFP⁺, CD95^highGFP⁺, CD95^highGL7⁺ lymph node cell sorted from immunized AID-Cre-YFP, AID-GFP⁺, and AID^−/− mice, respectively. Segments in pie charts are sized in proportion to the number of sequences carrying mutations as indicated. The number of sequences analyzed is given in the center of the pie charts. Mutation frequency and P values are indicated below. Statistical significance was assessed by a two-tailed t test assuming unequal variance and comparing values to mutation background determined in AID^−/− sequences.