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. 2007 Dec 26;28(5):1713–1723. doi: 10.1128/MCB.01360-07

FIG. 6.

FIG. 6.

p53−/− CAT-Tg DN lymphomas are immature pre-DP cells and are distinct from p53−/− DP/SP lymphomas. (A) Representative fluorescence-activated cell sorter profiles showing forward scatter (FSC) and side scatter (SSC) (top) and cell surface staining of CD4, CD8 (middle), and TCRβ (bottom) on electronically gated CD4− CD8− DN cells of total thymocytes from a thymic lymphoma-bearing 91-day-old p53−/− mouse. (B) Abundance of Deltex-1 and c-Myc mRNA relative to GAPDH in DN thymocytes from C56BL/6 and lymphoma-bearing p53−/− CAT-Tg mice analyzed by real-time PCR. (C) Total thymocytes from C57BL/6 and lymphoma-bearing p53−/−CAT-Tg mice were stained with a group of TCRα antibodies. (Top) Surface expression of TCRα on thymocytes electronically gated to exclude DP, SP, and NK cells. (Bottom) Surface expression of TCRα on electronically gated DP and SP thymocytes. (D) PCR analysis of rearranged TCRαV5H+F3+2C-J2T11 gene segments using the same amount of genomic DNA purified from thymocytes. (E) PCR analysis of rearranged TCRβD2-Jβ2 gene segments using same amount of genomic DNA purified from total thymocytes. (F) Chromosome structural aberrations in p53−/− CAT-Tg lymphomas. From left to right, a normal chromosome 6 (left), a deleted chromosome 6 (right), a chromatid break on chromosome 11, and chromosome translocations illustrated by T(10;17) (left) and T(4;6) (right) are shown; centromere fusions were sometimes simple, containing material from the same chromosome, as in Rb(3.3) on the left, and sometimes more complex, as in Rb[5.T(5;4)] on the right. Polycentric chromosomes resulted from the fusion of chromosomes in a tail-to-tail orientation, as in the dicentric chromosome on the left, or in a head-to-tail orientation, as occurred multiple times in the pentacentric chromosome on the right. (G) Spectral karyotype of p53−/− CAT-Tg lymphomas. The top metaphase is a pseudotetraploid cell (n = 80) with deletions of chromosome 5, a general loss of chromosome 13, and a gain of chromosome 17. Also illustrated at the bottom left side are two chromosome fragments containing centromeres (black arrows). The bottom cell is a pseudotriploid (n = 60) karyotype with a complex rearrangement between chromosomes 10 and 16 containing four centromeres (red arrows).