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Cytochemical evidence for the presence of actin in the nucleus of the voodoo lily appendix
Authors:Skubatz H  Orellana M V  Yablonka-Reuveni Z
Institution:(1) Department of Botany, University of Washington, Seattle, WA 98195, USA;(2) Department of Bioengineering, University of Washington, Seattle, WA 98195, USA;(3) Department of Biological Structure, University of Washington, Seattle, WA 98195, USA
Abstract:Immunoflorescence microscopy of sections of the voodoo lily Sauromatum guttatum appendix stained with monoclonal antibodies against agr-smooth muscle actin and cytoplasmic actin revealed different staining intensity of different parts of the cell. The anti-cytoplasmic-actin recognized antigens present mainly in the cytoplasm, and the anti-agr-smooth muscle-actin recognized more intensively antigens present in the nuclei. A positive staining of the nucleus was also obtained with FITC–phalloidin confirming the presence of actin in its filamenous form in the nucleus. The presence of a nuclear agr-smooth muscle-actin-like protein was further confirmed by confocal laser microscopy. On Western blots, the two anti-actins labelled a protein band that comigrated with standard actin at the approximate molecular weight of 43thinspkDa. Several other proteins interacted with the two antibodies to a different degree. The monoclonal antibodies against beta-tubulin subunit stained only the periphery of the cytoplasm and anti-pan cytoplasmic myosin stained the cytoplasm weakly. On a Western blot, anti-beta-tubulin subunit primarily recognized a protein band at the appropriate molecular weight of 50thinspkDa. This is the first cytochemical evidence for the presence of agr-smooth muscle-actin-like protein in the plant nucleus.
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