Fig 2. Effect of mir-511-3p target insertions on ZIKV replication in the serum, brain and testes of AG129 mice.

Adult AG129 male mice (n = 6–9 per group) were infected IP with 10⁶ pfu of ZIKV clones containing target sites for mir-511-3p [2×511(T)] or scrambled target sites [2×scr]. Mice were bled and/or sacrificed at the indicated days post infection. Mean viral titer ± SD in the serum (A), brain (B), or testes (C) was determined by titration in Vero cells. The dashed lines indicate the limit of virus detection: 1.5 log₁₀ pfu/mL of serum (A) and 1.7 log₁₀ pfu/g of brain or testis (B-C). Differences between the virus titers were compared using two-way ANOVA (**** p < 0.0001; *** p< 0.001; * p<0.05; ns—denotes not significant, p> 0.05). Except 6 dpi, all data for the 2×scr virus are retrospective [17] and provided here for comparison. Data for 6 dpi for the 2×scr virus was combined from the data reported earlier (n = 6) [17] and newly generated results (n = 3). Viral RNA was isolated from the serum at 1 dpi, brain at 12 dpi and testes at 12 dpi, and the region containing miRNA targets was sequenced. If mutation/deletion was detected in the miRNA targeted region, viral titer for these samples was reported under the name 2×511(T)/mut.