FIG. 3.

RASCAL intracellular localization. Mock-infected (A to C) or Towne/GFP-IE2-infected HF (MOI of 5) (D to R) were harvested at the indicated times p.i. and were stained with affinity-purified rabbit anti-RASCAL polyclonal Abs followed by Alexa Fluor 594-conjugated goat anti-rabbit Abs. The signal emitted from the GFP-IE2 protein was further amplified with FITC-conjugated anti-IE1/IE2 Abs, and nuclear DNA was stained with Hoechst 33342. The arrows point at RASCAL accumulation at the nuclear rim, the arrowheads indicate the peculiar structures observed on the nuclear surface at late times p.i., and the asterisks mark the locations of the cytoplasmic RASCAL-positive vesicles. Original magnification, ×400.