FIG. 2.

293Rep cells contain the HCV subgenomic replicon. (A) ³³P metabolic labeling of replicon RNA in the presence of actinomycin D. Total cellular RNA was extracted from cells treated with ³³P and actinomycin D for 16 h and was resolved in a 0.6% denaturing agarose gel. (B) Northern blots detecting the negative strand of HCV subgenomic replicons. Total cellular RNA was extracted and resolved in a glyoxal-dimethyl sulfoxide agarose gel. The RNAs were transferred and hybridized with an HCV negative-strand-specific ³²P-labeled RNA probe. The first three lanes are positive controls of in vitro-synthesized negative strand RNA, with 10⁸, 10⁷, and 10⁶ copies used, as indicated.