(A) Southern blot analysis to examine IgH gene configuration with an IgH-specific probe

Genomic DNA was prepared and digested with EcoRI from splenic cells of LMP1/CD40 mice that developed neoplasias (34, 142, 176, 197, 230, 247, 365, and 221), one young LMP1/CD40 mouse (900; 3 mo of age), as well as a control mouse (905). The sequence analysis of the amplified IgH genes is shown in Fig. S6. (B) Representative histological analyses of diseased LMP1/CD40 mice and age matched control mice. (1) HE-stained normal spleen. (2) HE-stained spleen representative for diseased LMP1/CD40 mice, showing prominent nodular tumor infiltrates. (3) Lymphoma infiltrate within the spleen of a representative diseased LMP1/CD40 mouse. B cells were visualized by immunohistochemistry using an antibody specific for B220. (4) Higher magnification of section described in 3. (5) Dispersed reactive T cells within the lymphoma infiltrate were detected by an anti-CD3–specific antibody. A representative highly magnified section is shown. Bars: (1 and 2) 400 μm; (3) 100 μm; (4, 5, and inset) 25 μm. (C) Flow cytometric analysis of splenic cells for the expression of IgM and IgD. The gates were set according to the age-matched controls. For the diseased LMP1/CD40 mice, a shift toward one or two main populations could be observed, as shown for three representative samples. Fig. S6 is available at .<b>Copyright information:</b>Taken from "Constitutive CD40 signaling in B cells selectively activates the noncanonical NF-κB pathway and promotes lymphomagenesis"The Journal of Experimental Medicine 2008;205(6):1317-1329.Published online 9 Jun 2008PMCID:PMC2413030.