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. 2007 Dec 24;204(13):3195–3208. doi: 10.1084/jem.20071224

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Copyright © 2007, The Rockefeller University Press

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Figure 3. — V_H cRS cleavage is detected only in pro–B cells from RAG2:GFP knock-in mice (21). (A) Sorted pro-, pre, and immature B cells were analyzed by RT- and LM-PCR. Mature, recirculating B cells (+IgD) were removed by incubation with anti-IgD (-IgD). RAG2:GFP fluorescence was detected in pro- and pre–B cells, but not in immature B cells. (B) RT-PCR for Rag1, Tdt, and GAPDH transcripts revealed RAG1 message in both pro–B and pre–B cells; Tdt expression was detected only in sorted pro–B cells. (C) LM-PCR for primary J_H, V_H, Jκ, and Dβ rearrangements demonstrate the lineage and developmental restriction of V_H cRS SEs, and confirms the purity of the sorted cell populations. V_H1, V_H2, and V_H5 cRS SEs were detected only in pro–B cells. CD14 PCR demonstrated the equivalence of genomic template.