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Subcellular Localization of an Oncoprotein using Immunofluorescence, Confocal Imaging and Immuno Electron Microscopy

Published online by Cambridge University Press:  02 July 2020

K. Murti ,
C. Caslini ,
P.H. Domer ,
S. J. Korsmeyer ,
J. Boer ,
G. Grosveld  and
A.T. Look
K. Murti
Affiliation:
St. Jude Children's Research Hospital, Memphis, TN38105-2794
C. Caslini
Affiliation:
St. Jude Children's Research Hospital, Memphis, TN38105-2794
P.H. Domer
Affiliation:
Washington University School of Medicine, St. Louis, MO63110
S. J. Korsmeyer
Affiliation:
Washington University School of Medicine, St. Louis, MO63110
J. Boer
Affiliation:
St. Jude Children's Research Hospital, Memphis, TN38105-2794
G. Grosveld
Affiliation:
St. Jude Children's Research Hospital, Memphis, TN38105-2794
A.T. Look
Affiliation:
St. Jude Children's Research Hospital, Memphis, TN38105-2794
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Extract

The mixed-lineage leukemia (MLL) gene is associated with more than 25 translocations of chromosome band 1 lq23 in the human leukemias. In certain unusually aggressive leukemias, the N-terminal portion of MLL fuses in frame to a portion of the transcription factor AF4. As a part of our investigation of the role of the MLL-AF4 fusion oncoprotein in Leukemogenesis, we studied the cellular sites of distribution of MLL-AF4 in the Hela carcinoma cells and U937 myeloid cells by immunofluorscene, confocal imaging and immuno-electron microscopy. Hela and U937 cells were stably transfected with a FLAG epitope-tagged MLL-AF4 (F-MLL-AF4) under the control of a tetracycline-responsive promoter and the expression of F-MLL-AF4 detected by indirect immunofluorescence (IF) with an anti-FLAG monoclonal antibody. These studies revealed that in both cell types the antibodies labeled two nuclear components, the nucleoli in an intense manner and the nucleoplasm in a punctate (100 to 200 dots) pattern. No antibody staining was observed in control (in the presence of tetracycline) cells.

Type
Cytochemistry (Light and Electron Histochemistry)
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 1096 - 1097
Copyright
Copyright © Microscopy Society of America

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