Effect of Tudor-SN knockdown on LGTV infection.

(A) I. scapularis IDE8 cells were treated with Tudor-SN, Rs86 or GFP control dsRNAs and infected with LGTV at MOI 0.1. LGTV RNA levels were determined after 24h and 48h by NS5 real-time PCR normalizing against tick β-actin mRNA levels and are presented relative to the GFP dsRNA control. Relative LGTV RNA levels were compared between groups by two-way ANOVA and were not statistically different between Tudor-SN and GFP dsRNA-treated cells (P>0.05; N = 8). (B) I. scapularis IDE8 cells were treated with Tudor-SN, Rs86 or GFP control dsRNAs and infected with LGTV at MOI 0.1. Virus titers in the cell supernatants were determined by plaque assay after 24h and 48h and are presented relative to the GFP dsRNA control. Relative LGTV titers were compared between groups by two-way ANOVA and were not statistically different between Tudor-SN and GFP dsRNA-treated cells (P>0.05; N = 8). (C) I. ricinus IRE/CTVM20 cells were treated with Tudor-SN, Rs86 or GFP control dsRNAs and infected with LGTV at MOI 0.1. LGTV RNA levels were determined after 24h and 48h by NS5 real-time PCR normalizing against tick β-actin mRNA levels and are presented relative to the GFP dsRNA control. Relative LGTV RNA levels were compared between groups by two-way ANOVA (P≤0.05; N = 8). (D) I. ricinus IRE/CTVM20 cells were treated with Tudor-SN, Rs86 or GFP control dsRNAs and infected with LGTV at MOI 0.1. Virus titers in the cell supernatants were determined by plaque assay after 24h and 48h and are presented relative to the GFP dsRNA control. Relative LGTV titers were compared between groups by two-way ANOVA and were not statistically different between Tudor-SN and GFP dsRNA-treated cells (P>0.05; N = 8). For each tick cell line, two independent experiments with 4 replicates each were conducted and combined after normalization.