Fig. 7. Effect of sfRNA deficiency on replication of ISFs in mosquito cells.

A Northern blot analysis of sfRNA production by WT and PK mutants of PARV, PCV and BinJV. C6/36 cells were infected with WT or mutant viruses at MOI = 1. Total RNA was isolated at 7 dpi and used for blotting. The bottom panel show Et-Br-stained 5.8 S rRNA as an equal loading control. Images are representative of two independent experiments that produced similar results. PK1’, PK2, and PK3’ mutations represented substitutions of PK-forming nucleotides in L2 loops of SLI, SLII and SLII, respectively, with complementary nucleotides and are the same as in Fig. 2D (BinJV), 3 C (PaRV), 3 G (PCV). B Virus titers in culture fluids of RML-12 cells infected with WT or PK-mutants of ISFs. Cells were infected at MOI = 0.1, culture fluids were sampled at the indicated time point, and titers were determined using foci-forming immunoassay on C6/36 cells. Values are the means from three biological replicates ± SD. Statistical analysis was performed by two-way ANOVA with Dunnett’s correction, all comparisons were to WT, test was two-sided.