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. 2006 Sep 4;203(9):2085–2094. doi: 10.1084/jem.20061067

Figure 4.

Figure 4.

Cytokine induction of switch-associated mutations in LPS-cultured B cells. (A) In vitro class switching of splenic B cells from a wild-type (control) C57BL/6 mouse analyzed by flow cytometry after a 7-d culture with LPS plus either IL-4 or IFN-γ. The numbers indicate the percentage of events in the top right quadrants. Results are representative of two independent experiments. (B) Germline transcription in the in vitro–stimulated splenic B cells. Germline transcripts comprising IH and CH exons of each isotype were detected by RT-PCR from RNA extracted at day 3 of culture. hprt, hypoxanthine phosphoribosyltransferase control. (C) Distribution of transitions at C:G pairs in genomic DNA purified from splenic B cells from msh2 −/− ung −/− mice after a 7-d culture with LPS plus IL-4 or IFN-γ as indicated. For each isotype, a region that had been demonstrated to be targeted by AID in vivo (Fig. 2) was amplified and sequenced. The number of sequenced for each region vary as a consequence of sequencing failures. Data shown are from two independent experiments.