Telomeric IGH Losses Detectable by Fluorescence in Situ Hybridization in Chronic Lymphocytic Leukemia Reflect Somatic VH Recombination Events Iwona Wlodarska,

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Telomeric IGH Losses Detectable by Fluorescence in Situ Hybridization in Chronic Lymphocytic Leukemia Reflect Somatic VH Recombination Events Iwona Wlodarska, Christine Matthews, Ellen Veyt, Helena Pospisilova, Mark A. Catherwood, Tim S. Poulsen, Vera Vanhentenrijk, Rachel Ibbotson, Peter Vandenberghe, T.C.M. “Curly” Morris, H. Denis Alexander The Journal of Molecular Diagnostics Volume 9, Issue 1, Pages 47-54 (February 2007) DOI: 10.2353/jmoldx.2007.060088 Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Examples of FISH results. Analyzed cases: case 5 (A–C, F), case 1 (D), case 3 (E), diffuse large B cell lymphoma with t(3;14)(q27;q32) and +der(3)t(3;14)(q27;q32) (G) and normal peripheral B cells analyzed by FICTION with CD20 (H). Applied probes: LSI IGH/CCND1 (A–C), LSI IGH (D–F), 965B13-SO/47P23-SG (G), and 47P23-SG/2548B8-SO (H). Arrow and arrowheads in G indicate normal chromosome 14 and two der(3)t(3;14), respectively. Extra red signals seen in G correspond to 15q11, a region of co-hybridization of 965B13.9 The Journal of Molecular Diagnostics 2007 9, 47-54DOI: (10.2353/jmoldx.2007.060088) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 FISH mapping of IGH deletions in 11 CLL cases. Schematic presentation of the IGH region (top panel), applied probes (center panel), and FISH results (bottom panel) (not to scale). Checkered and dotted bars indicate Spectrum Green- and Spectrum Orange-labeled probes, respectively. Open bars mark regions found to be present on both chromosomes 14; filled bars point sequences lost on one homolog 14. The Journal of Molecular Diagnostics 2007 9, 47-54DOI: (10.2353/jmoldx.2007.060088) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Decision trees to identify IGH aberrations not specific for the applied FISH assay observed in B cell malignancies analyzed with either LSI IGH/CCND1 (A) or LSI IGH/CCND1 XT, -/BCL2, and -/CMYC (B). The latter assay is combined with the control Spectrum Aqua CEP 8, which is not included in this scheme. Cells harboring biallelic VH deletions are marked by an asterisk. The Journal of Molecular Diagnostics 2007 9, 47-54DOI: (10.2353/jmoldx.2007.060088) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions