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. 2004 May;78(9):4684–4699. doi: 10.1128/JVI.78.9.4684-4699.2004

FIG. 2.

FIG. 2.

Effect of the D215N vhs mutation on the RNase nuclease activity of yeast extracts. (A) Nuclease activity of cell extracts. Internally labeled pCITE-1 RNA was added to extracts of yeast harboring the empty expression vector (EV), 2.1 vhs expression vector (2.1vhs), or 2.1 expression vector bearing the D215N mutation (D215N), either alone or supplemented with RRL. In addition, a portion of the RNA was incubated in RRL (RRL). Samples were withdrawn at the indicated times, and RNA was extracted and analyzed as described in the legend for Fig. 1. (B) The data obtained in the experiment shown in panel A were quantified by phosphorimager analysis. (C) Western blot analysis of yeast extracts. Aliquots of the extracts used for panel A (each containing 20 μg of protein) were subjected to electrophoresis through an SDS-10% polyacrylamide gel, transferred to a nitrocellulose membrane, and then analyzed by Western blotting using a polyclonal rabbit anti-vhs antiserum (AE328).