Journal: Journal of Virology
Article Title: The Simian Virus 40 Late Viral Protein VP4 Disrupts the Nuclear Envelope for Viral Release
Figure Lengend Snippet: VP4 is concentrated along the nuclear envelope. (A) Cos7 cells were transiently transfected with either empty vector (−) or VP4 (+) in the presence of the tetracycline repressor. Tetracycline was added for the indicated times. Whole-cell lysates were immunoblotted with antisera against myc to detect VP4 or the loading control GAPDH. (B) Confocal immunofluorescence microscopy localization of VP4 in Cos7 cells was determined after 6 h of induction. Samples were labeled with myc (VP4-myc), ERp57, emerin, or lamin antisera and stained with either Hoechst or DAPI (DAPI, 4′,6-diamidino-2-phenylindole) as indicated. A fraction of the VP4-expressing cells displayed profound nuclear envelope disruption, as displayed in the lamin lower panel. Bars, 10 μm.
Article Snippet: The following antibodies were obtained as indicated: myc, lamin A/C, and emerin (Cell Signaling, Danvers, MA); GAPDH (glyceraldehyde-3-phosphate dehydrogenase; Millipore, Billerica, MA); VP1 and VP2/3 (Abcam, Cambridge, MA; VP2/3 was also obtained from A. Oppenheim, Jerusalem, Israel); calnexin (Enzo Life Sciences Inc., Ann Arbor, MI); large T antigen (LT; Merck KGaA, Darmstadt, Germany); and glutathione S -transferase (GST; Abmart, Arlington, MA).
Techniques: Transfection, Plasmid Preparation, Immunofluorescence, Microscopy, Labeling, Staining, Expressing