FIG 6.

Effect of spliceostatin A on segment 7 mRNA localization. (A) 293T cells were infected with reverse genetics PR8 virus at an MOI of 5 and at 6 h p.i. treated or mock treated with 100 μg/ml spliceostatin A. At 6 h p.i. samples were fixed and stained by FISH for segment 7 mRNA (red) or by 4′,6′-diamidino-2-phenylindole for DNA (blue) and imaged by confocal microscopy. Single optical slices are shown. Scale bar, 10 μm. (B) Cell lysates were examined by Western blotting for the indicated polypeptides. (C) Total cellular RNA was analyzed by radioactive primer extension, urea-PAGE, and autoradiography for the indicated RNA species.