Tudor-SN expression in response to pathogen infection.

(A) Differential expression/representation of Tudor-SN and Ago genes/proteins in I. scapularis nymphs, adult female guts and salivary glands in response to infection with A. phagocytophilum. Data was obtained from transcriptomics and proteomics analyses and values are shown as infected/uninfected Log2-fold ratio (P<0.05). Abbreviations:-, down-regulated/under-represented in infected ticks; +, up-regulated/over-represented in infected ticks; NS, no significant difference between infected and uninfected ticks; NF, not found in trascriptomics or proteomics data. (B) Tudor-SN mRNA levels in I. scapularis ISE6 cells in response to early (day 6) and late (day 13) infections with A. phagocytophilum. Five independent experiments were conducted for each early and late infection. Tudor-SN mRNA levels were determined by real-time RT-PCR in uninfected and infected cells and the infected-to-uninfected ratio of Ct values normalized against tick 16S rRNA and cyclophilin are shown in arbitrary units (Ave+SD). Normalized Tudor-SN mRNA levels were compared by Student’s t-test with unequal variance and were not statistically different between infected and uninfected cells (P = 0.05; N = 5). (C) Tudor-SN mRNA levels in I. ricinus ticks during TBEV infection. Tudor-SN mRNA levels were determined by real-time RT-PCR in the guts and salivary glands of female ticks (N = 10 ticks for each time point) uninfected and artificially infected with TBEV and fed on mice for 0, 1, 3 and 5 days. Tudor-SN Ct values normalized against tick 16S rRNA are shown in arbitrary units (Ave±SD) and were used to calculate infected/uninfected ratios and compared between Days 1–5 and Day 0 by Student’s t-test with unequal variance (*P<0.05; N = 10).