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N. Scott Reading, Stephen D. Jenson, Jeffrey K. Smith, Megan S

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1 5′-(RACE) Identification of Rare ALK Fusion Partner in Anaplastic Large Cell Lymphoma 
N. Scott Reading, Stephen D. Jenson, Jeffrey K. Smith, Megan S. Lim, Kojo S.J. Elenitoba-Johnson  The Journal of Molecular Diagnostics  Volume 5, Issue 2, Pages (May 2003) DOI: /S (10) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 Histology of the axillary lymph node biopsy showing lymphohistiocytic variant of ALCL. A: H&E stain showing complete effacement of nodal architecture by a proliferation of lymphoid cells with large pleomorphic folded nuclei with visible nucleoli and abundant eosinophilic cytoplasm. B: The tumor cells show strong positive reactivity for CD30 in a characteristic membrane and Golgi pattern. C: The tumor cells show uniform reactivity for ALK in an exclusively cytoplasmic and membrane distribution. The Journal of Molecular Diagnostics 2003 5, DOI: ( /S (10) ) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Analysis of 5′-RLM-RACE and cloning of variant ALK fusion transcript. A: Lane 1, reverse transcriptase-nested PCR products of 5′-RLM-RACE, arrow indicates the selected band for cloning; lane 2, linear DNA size markers. B: ALK-specific PCR analysis of selected plasmids containing the ∼1000-bp 5′-RLM-RACE product. Arrow indicates the ALK PCR product. Lane 1, template-free control (H2O); lane 2, clone 1; lane 3, clone 2; lane 4, clone 3; lane 5, clone 4; lane 6, linear DNA size markers. The Journal of Molecular Diagnostics 2003 5, DOI: ( /S (10) ) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

4 Figure 3 Schematic and fusion point sequence of cloned variant ALK fusion gene. The TPM3-ALK fusion transcript contained TPM3 5′-nontranslated sequence (crosshatch) and coding sequence (white), and partial ALK sequence (diagonal slash). The DNA sequence of the variant TPM3-ALK transcript fusion site is outlined below. The predicted amino acid sequence is also shown. As indicated by the vertical line the TPM3-derived portion of the fusion transcript ends at nucleotide 1 of codon 222 and the portion derived from ALK begins with nucleotide 2 of codon 1058. The Journal of Molecular Diagnostics 2003 5, DOI: ( /S (10) ) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

5 Figure 4 Analysis of T-cell clonality by PCR. Lane 1, template-free control (H2O); lane 2, negative (polyclonal) DNA control from a hyperplastic tonsil; lane 3, positive (monoclonal) DNA control from the Jurkat T-cell lymphoma cell line; lanes 4 and 5, monoclonal TCR-γ bands of identical size in duplicate reactions of patient's biopsy specimen; lane 6, DNA size marker. The Journal of Molecular Diagnostics 2003 5, DOI: ( /S (10) ) Copyright © 2003 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions


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