An anti-Bcl-2 antibody prevents 2-deoxy-D-ribose-induced apoptosis in the IPLB-LdFB insect cell line |
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Authors: | D Barbieri D Malagoli B Cuoghi E Ottaviani |
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Institution: | (1) Department of Animal Biology, University of Modena and Reggio Emilia, via Campi 213/D, 41100 Modena (Italy), Fax: +39 059 205 55 48; e-mail: ottaviani.enzo@unimo.it, IT |
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Abstract: | Confocal microscopy reveals that the anti-Bcl-2 antibody (pAb) is able to diffuse across the plasma membrane of the fat body
cell line IPLB-LdFB from the insect Lymantria dispar, demonstrating the presence of Bcl-2-like molecules in the cytoplasm. Immunoperoxidase procedure confirms the cellular localization.
Furthermore, an immunoprecipitation corresponding to a molecular weight of 29 KDa is observed with western blot analysis using
the anti-Bcl-2 pAb. Cytofluorimetric experiments show that anti-Bcl-2 pAb counteracts 2-deoxy-d-ribose-induced apoptosis and provokes morphological changes in the insect cell line, i.e. a reduction in cell size, the disappearance
of the vacuola and changes in shape. At the same time, the antibody provokes mitochondrial membrane depolarization, and N-acetyl-l-cysteine is unable to reconstitute the physiological conditions. The present findings suggest that Bcl-2-like proteins play
a main role in maintaining of the integrity of cellular components, e.g. mitochondria, rather than in controlling programmed
cell death.
Received 23 January 2001; received after revision 1 March 2001; accepted 1 March 2001 |
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Keywords: | , IPLB-LdFB insect cell line, anti-Bcl-2 antibody, 2-deoxy-D-ribose, N-acetyl-l-cysteine, valinomycin, apoptosis,,,,,,mitochondrial membrane potential, confocal scanning laser microscopy, immunocytochemistry, western blot, flow cytometry, |
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