1. Detection of Normal and Chimeric Nucleophosmin in Human Cells
by Jacqueline L. Cordell, Karen A.F. Pulford, Barbara Bigerna, Giovanna Roncador, Alison Banham, Emanuela Colombo, Pier-Giuseppe Pelicci, David Y. Mason, and Brunangelo Falini
Blood
Volume 93(2):
January 15, 1999
©1999 by American Society of Hematology

2. Jacqueline L. Cordell et al. Blood 1999;93:632-642
©1999 by American Society of Hematology

3. Jacqueline L. Cordell et al. Blood 1999;93:632-642
©1999 by American Society of Hematology

4. Immunostaining of cytocentrifuged cell lines and paraffin-embedded normal tissues for NPM (APAAP technique).
Immunostaining of cytocentrifuged cell lines and paraffin-embedded normal tissues for NPM (APAAP technique). Cell lines: The t(2;5)-positive cell line Karpas 299 (a) and a murine mastocytoma cell line (b) both show labeling of nucleoli with antibody to the C-terminal portion of NPM (antibody NPMc). Note the cytoplasmic labeling of a mitotic cell in (b). Kidney: (a) Many cells show nuclear labeling for nucleophosmin. (b) and (c) show high power views of, respectively, a glomerulus and tubular epithelium (antibody NA24). Thymus: Two different anti-NPM antibodies give essentially identical labeling patterns of cell nuclei in a Hassall’s corpuscle and in the surrounding medullary tissue. Tonsil: Nuclear labeling of lymphoid cells is seen. The arrows in the low power view (left) indicate scattered mitotic cells in which there is cytoplasmic labeling, seen at high power on the right (antibody NA24). Cerebral cortex: Neural cells show strong cytoplasmic staining, seen at low power (above) and high power (below) (antibody NA24).
Jacqueline L. Cordell et al. Blood 1999;93:
©1999 by American Society of Hematology

5. In a squamous cell carcinoma, the nuclei of all cells are labeled (antibody NA24 APAAP technique).
In a squamous cell carcinoma, the nuclei of all cells are labeled (antibody NA24 APAAP technique). Occasional mitotic cells (arrows) show diffuse cytoplasmic labeling.
Jacqueline L. Cordell et al. Blood 1999;93:
©1999 by American Society of Hematology

6. Staining of paraffin sections of an ALK-positive ALCL (APAAP technique).
Staining of paraffin sections of an ALK-positive ALCL (APAAP technique). (Upper) The reactivity of the same area of lymph node with antibodies to the C-terminal epitope (antibody NPMc) and to the N-terminal region (antibody NA24) is seen at low power (above) and high power (below). The tumor cells surround a residual area of normal B cells (asterisks). A clear difference between the labeling patterns of the two antibodies is seen even at low magnification, accounted for by the reactivity of the anti–N-terminal antibody with tumor cell cytoplasm. Scattered mitotic cells showing diffuse cytoplasmic labeling are seen in both sections (arrowed) and can be picked out in the low-power view of the immunostaining for the C-terminus. (Lower) The appearance of the tumor in sections stained by hematoxylin and eosin and for ALK protein is shown at low power (left) and high power (right).
Jacqueline L. Cordell et al. Blood 1999;93:
©1999 by American Society of Hematology

7. Staining of paraffin sections of a second ALK-positive ALCL seen at low (left) and high (right) power (APAAP technique).
Staining of paraffin sections of a second ALK-positive ALCL seen at low (left) and high (right) power (APAAP technique). As in the case shown in Fig 4, a clear difference is seen between the immunostaining for the C-terminal epitope (antibody NPMc) and an N-terminal epitope (antibody NPMa). Immunostaining for ALK is also shown. In the lower power view, the corresponding areas in three sections are shown. The asterisks indicate residual normal lymphoid tissue surrounded by tumor cells.
Jacqueline L. Cordell et al. Blood 1999;93:
©1999 by American Society of Hematology

8. Schematic diagram of NPM and ALK proteins in normal cells and in t(2;5)-positive lymphoma cells and the resulting immunostaining patterns.
Schematic diagram of NPM and ALK proteins in normal cells and in t(2;5)-positive lymphoma cells and the resulting immunostaining patterns. This scheme assumes that NPM and NPM-ALK are present in both monomeric and dimeric forms in approximately equal numbers, and does not show larger oligomers that might form.5 It may thus represent an oversimplification of what occurs in vivo. (a) In a normal cell, NPM is confined to the nucleus. In a cell carrying the (2;5) translocation, NPM-ALK is found in the cytoplasm and other NPM-ALK that has heterodimerized with wild-type NPM has acquired nuclear localization. (b) Immunostaining for N- and C-terminal epitopes of NPM and for ALK show distinctive patterns, reflecting the localization of the proteins in the scheme shown above.
Jacqueline L. Cordell et al. Blood 1999;93:
©1999 by American Society of Hematology