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. 2016 Jun 14;7:11889. doi: 10.1038/ncomms11889

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(a) Partial G-banded karyotype showing a t(10;14)(q24;q32) translocation in a patient with SMZL (case 1). Arrows mark the derivative chromosomes 10 and 14. (b,d) Interphase FISH analysis of bone marrow cells from two patients with t(10;14) using an NKX2-3 break-apart assay. Cells carrying the translocation show split of green and red probes (green arrows), in addition to the co-localized signals on the normal allele (red arrows). Scale represents 2 μm in all cases. (c) Ideogram depicts location of breakpoints cloned by LDI-PCR from the IGHS segment in the t(10;14)(q24;q32) from case 1. Representative breakpoint sequences with identity to IGHS and the corresponding NKX2-3 gene are shown. (e) Expression of NKX2-3 by reverse transcription–PCR cell subpopulations in peripheral blood of healthy donors. (f) NKX2-3 expression measured by quantitative RT–PCR in samples obtained from patients with mature B-cell malignancies and in non-tumoral cells isolated from healthy donors: CD19⁺ B cells, CD3⁺ T cells and CD14⁺ myeloid cells (peripheral blood); pre-germinal centre IgD⁺ naïve B cells, post-germinal center CD27⁺ memory B cells and CD71⁺ germinal centre centroblasts (tonsils); and CD34⁺ haematopoietic stem/progenitor cells (bone marrow). The cutoff value for positive NKX2-3 expression was considered when greater than s.d. (standard deviation) x4 of the mean value of the expression of CD19⁺ cells plus CD34⁺ cells. B-CLL, B-cell chronic lymphocytic leukaemia; DLBCL, diffuse large B-cell lymphoma; FCL, follicular lymphoma; MALT, mucosa-associated lymphoid tissue lymphoma; MCL, mantle cell lymphoma; MM, multiple myeloma; SMZL, splenic marginal-zone lymphoma. The number of patient samples analysed in each experiment is shown. (g) IHC analysis of non-tumoral human spleen tissues using moAbs for NKX2-3 (clone 454C/H9), CD34 and CD68. (h) IHC analysis of different tissues samples from the case with t(10;14)(q24;q32) (case 1). In the left panel, the bone marrow biopsy obtained at diagnosis shows expression of NKX2-3 in scattered lymphoid B cells. In the middle and right panels, sequential lymph node biopsies obtained during histological transformation to non-GC DLBCL and subsequent relapse display nuclear expression of NKX2-3 in large B-cell lymphoma cells. Scale represents 100 μm in all cases.