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. 1998 Dec 21;188(12):2369–2374. doi: 10.1084/jem.188.12.2369

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Flow cytometric analysis of LPS-stimulated splenic B cells of pooled homozygous targeted mice and their wild-type littermates. By depletion of CD43⁺ spleen cells using the MACS system, resting B cells (CD43⁻) were isolated and cultured with LPS or with LPS + IL-4. After 5 d cells were fixed and stained intracellularly for expression of IgG1. Due to constitutive activity of the hMT promoter, class switch to IgG1 is independent of IL-4 in B cells from targeted mice. A representative example of two independent experiments is shown. (Note that in all cultures with IgG1 switched B cells, there is a basal level staining of all B cells for IgG1, probably due to Fc-γR bound IgG1 on the surface.)