Verification of Cct1 and Cct2 roles in modulating FHV RNA replication in replicon-bearing S2 cells. (A) Northern blot of FHV genomic (+)RNA1 and subgenomic (+)RNA3 accumulation in FHV replicon-bearing S2 cells after RNAi-mediated knockdown of Cct1, Cct2, or both. Mock transfected cells (lane 1) or cells treated with dsRNA against LacZ (lane 2), FHV RNA1 (lanes 3), Cct1 (lane 4), Cct2 (lane 5), or both Cct1 and Cct2 (lane 5) and co-transfected with pS2F1 and pS2LacZ were induced with 0.5 mM Cu2+ for 18 h and FHV RNA accumulation was determined by northern blotting as described in Fig. 5. The position of FHV subgenomic (+)RNA3 is shown on the right, and ribosomal RNA (rRNA) bands are shown as loading controls. The position of genomic (+)RNA1, which is barely detectable in transiently transfected S2 cells, is also shown on the right. (B) Quantitative data for subgenomic (+)RNA3 and β-galactosidase activity in S2 cells co-transfected with pS2F1 and pS2LacZ after RNAi-mediated knockdown of the indicated dsRNA targets. P-value < 0.05* or 0.005**.