Figure 2.

Enrichment of ssRNA and dsRNA Fractions of CVB3-Infected Cells

(A) Viral RNA species produced during picornavirus infection. RF, replicative form. RI, replication intermediate. Purple line, (+)RNA; blue line, (−)RNA; orange circle, VPg.

(B) HeLa cells were infected with CVB3 in the absence or presence of replication inhibitor GuHCl (2 mM) for 5 hr and total RNA was extracted. ssRNA and dsRNA fractions were separated by LiCl differential precipitation and examined on agarose gel. Note that the ssRNA fraction was diluted 400-fold relative to the dsRNA fraction to load comparable amounts of RNAs. M, dsDNA marker with indicated size in kbp; closed arrow, viral ssRNA; arrowhead, viral dsRNA; asterisk, unknown bands.

(C) vRNA purified from pelleted virus particles was visualized on agarose gel next to the same DNA marker as used in (A).

(D) HeLa cells were infected with CVB3 (MOI 100) in the absence or presence of GuHCl (2 mM) for 2.5 hr and the dsRNA fraction was purified from total RNA extract by LiCl precipitation and examined on gel. M, dsDNA marker; arrow, viral dsRNA; asterisk, unknown bands.

See also Figure S2.