FIG. 4.

Alterations to the SUP locus affects the steady-state accumulation of multiple RNA species. U373 cells were either infected with each individual virus at high MOI or mock infected in the presence (+) or absence of drug (CHX or PAA). At 6 h postinfection, total RNA was harvested, fractionated by electrophoresis through formaldehyde-agarose gels, and transferred to nylon membranes. To detect transcripts that span the Us-TRs junction, strand-specific RNA probes were prepared from the BstE2-XbaI region (see map in Fig. 1). RNAs were detected only with probes antisense to the previously characterized Us11 and Us12 ORFs; furthermore, identical RNAs were observed in WT virus- and Δ34.5-infected cells when an antisense probe within the SUP1 deletion (ApaLI-EcoNI) was employed (not shown). Approximate sizes of the RNA species (shown to the right of the gel) are as follows: a (Us11), 1.4 kb; b (Us12), 1.9 kb; c, 2.6 kb; d, 3.4 kb; e, 7.9 kb.