Open Access Highly Accessed Research article

Identification of a novel Rev-interacting cellular protein

Susanne Kramer-Hämmerle1, Francesca Ceccherini-Silberstein12, Christian Bickel1, Horst Wolff1, Michelle Vincendeau1, Thomas Werner3, Volker Erfle1 and Ruth Brack-Werner1*

Author Affiliations

1 Institute of Molecular Virology, GSF-National Research Center for Environment and Health, Ingolstädter Landstraße 1, D-85764 Neuherberg, Germany

2 Department of Experimental Medicine, University of Rome Tor Vergata, Via Montpellier 1, Rome 00133, Italy

3 Genomatix Software GmbH, Landsbergerstr. 6, D-80339 München, Germany

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BMC Cell Biology 2005, 6:20  doi:10.1186/1471-2121-6-20

Published: 24 April 2005

Additional files

Additional File 1:

Schematic representation of cDNAs with 16.4.1-coding sequences in predicted open reading frames. The scheme shows various cDNAs with 16.4.1 sequences identified by BLAST search of Entrez databases ([73] (indicated by lines). Open reading frames (ORF) were predicted with the ATGpr program [77]. Bars indicate the locations of the predicted ORFs within the cDNAs. The positions of the potential translation initiation and stop codons are marked by M and *, respectively. Regions with 16.4.1 encoding sequences are labeled yellow. All accession numbers are from the GenBank database.

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Additional File 2:

Sizes of hypothetical 16.4.1 cDNAs and proteins. The table indicates the lengths of the cDNAs shown in Additional file 1: Figure A1 and the calculated sizes of the proteins they are predicted to encode.

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Additional File 3:

Analysis of expression of 16.4.1 proteins with a monoclonal antibody directed against 16.4.1. Figure A2 shows expression of 16.4.1 proteins in HeLa cells. (A) Immunohistochemical analysis. Expression of 16.4.1 was detected in HeLa cells transfected with pIgG-16.4.1 (panel a) and in non-transfected HeLa cells (panel b) by indirect immunofluorescence with the monoclonal antibody against 1.6.4.1 and Cy3-labelled secondary antibodies. Panel c shows lack of reactivity of non-transfected HeLa cells with the secondary antibody. All images were taken with the same exposure times (300 ms). (B) Western blot analyses. Additional to 16.4.1-GFP several proteins are detected in HeLa cells expressing 16.4.1-GFP after transfection of pC16.4.1sg143. Proteins in whole-cell lysates were separated by electrophoresis in gradient gels (4–12% and 3–8%), and blotted onto nitrocellulose membranes. 16.4.1 proteins were detected with the monoclonal 16.4.1 antibody and a secondary antibody conjugated with horse radish peroxidase (HRP). For detection of the 16.4.1-GFP fusion protein, the membranes were stripped and reprobed with polyclonal antibodies against GFP. Table A2 lists proteins recognized by the monoclonal antibody against 16.4.1 in different human cell lines and primary human tissues by Western blot analyses. HeLa (cervix carcinoma) and 293T (embryonal kidney) cell lines are non-neural cells and U138MG, U251MG and 85HG66 human glioblastoma cell lines. Human cortical brain tissue and peripheral blood mononuclear cells were also investigated.

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