FIGURE 6.

Determination of molecular mass of HCV RNP complex. (A) The RNA in the HCV RNP complexes was ³²P-radiolabeled in the presence of [α-³²P]CTP by a cell-free replication assay as described previously (Ali et al. 2002). (B) The proteins in the HCV RNP complexes were metabolically labeled with ³⁵S-methionine. The RNP complexes derived from FCA4 or Huh7 cellular lysates were purified by avidin-agarose affinity purification (Fig. 1 ▶) and treated with 1% NP-40 for 20 min at 4°C and electrophoresed on 5% native PAGE at 100 V for 24 h at room temperature. RNP complexes were detected by autoradiography (A, lane 3; B, lane 2). (B, lane 1) Huh7 lysates were used as the control. (A, lanes 1,2) Molecular weight markers including apoferretin (443 kD) and thyroglobulin (669 kD; Sigma) were stained with Coomassie brilliant blue.