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. 2006 Oct 2;203(10):2305–2314. doi: 10.1084/jem.20060411

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Copyright © 2006, The Rockefeller University Press

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Figure 1. — Prdm1 gene deletion and phenotypic analysis of PerC and splenic B-1 cells. (A) Quantitative real-time PCR for prdm1 performed on genomic DNA from two purified CD19^Cre/+prdm1^Flox/Flox (open bars) and one littermate control (filled bar) PerC B-1 cell cultures. Primers were designed to amplify the floxed, but not the deleted, allele (see Materials and methods), and DNA loading was normalized to the peptidyl prolyl isomerase A gene. Percentages of prdm1^Flox/Flox amplified DNA are shown. (B) Representative flow cytometry analysis from one purified CD19^Cre/+ prdm1^Flox/Flox B-1 cell culture used for deletion analysis in A, stained with antibodies against IgM and Mac-1. (C) Total PerC cells from CD19^Cre/+ prdm1^Flox/Flox mice (right) and littermate control (left) mice stained with antibodies against B220 and CD5. Upper gate is B-1a, and the lower gate is B-1b. Mean ± SEM, n = 7. (D) Bar graph shows averages and SEM of total PerC B-1 cells (IgM⁺Mac-1⁺), n = 7. (E) Total splenocytes stained with antibodies for CD43, B220, and CD5. B220 and CD5 expression on CD43⁺ cells are shown. Mean ± SEM, n = 4.