Fig. 3

Validation of potential targeting relationship between miRNA-novel-53 and ModSP. A Sequence alignment of miRNA-novel-53 and the predicted binding site in ModSP. ModSP-WT, predicted target sites in ModSP; ModSP-MT, the binding sites of ModSP were mutated into noncomplementary bases with the miRNA-novel-53 seed sequence. B Relative luminescence activity after transfection of different plasmids in the S2 cell line. 53 + WT, cells transfected with pAc5.1b-miRNA-novel-53 and psiCHECK2-ModSP-WT plasmids; 53 + MT, cells transfected with pAc5.1b-miRNA-novel-53 and psiCHECK2-ModSP-MT plasmids; pAc + WT, cells transfected with pAc5.1b empty vector and psiCHECK2-ModSP-WT plasmid; pAc + pSi, cells transfected with pAc5.1b and psiCHECK2 plasmids. Each sample had in four replicates. Statistical differences between sample means were determined using the Student’s t-test. ns, P > 0.05; **P < 0.01. C Localization of miRNA-novel-53 and ModSP was detected with FISH in Aag2 cells. Green, ModSP; red, miRNA-novel-53; blue, Hoechst 33342. Scale bar, 10 μm. D Relative abundance of precipitated ModSP mRNA by Ae. aegypti AGO1 RIP in the different samples. **P < 0.01