Fig. 6: Vitellogenin is a direct target and is upregulated by hsa-miR-21-5p.

a Mosquitoes were fed with artificial diet only or on diet containing NC mimic or hsa-miR-21-5p mimic. They were collected 12 h after feeding, and vitellogenin levels relative to RPS17 were determined using RT-qPCR. Each data point represents a single mosquito. One-way ANOVA with Tukey’s post-hoc multiple comparisons were used for data analysis. ns, not significant; **p < 0.01. b Mosquitoes were injected with APS buffer, a negative control (NC) inhibitor or hsa-miR-21-5p inhibitor. Four days after injection mosquitoes were fed on a human volunteer and 12 h after feeding levels of vitellogenin relative to RPS17 were detected using RT-qPCR. One-way ANOVA with Tukey’s post-hoc multiple comparisons were used for data analysis. ns, not significant; *p < 0.05. c Bioinformatic analysis using RNAHybrid (v2.2.1) showing two different target sites in the vitellogenin gene. d The vitellogenin target sequences were cloned downstream of GFP in the pSLfa vector. e Aag2 cells were co-transfected with the plasmid and NC mimic, hsa-miR-21-5p mimic, or two different mutant mimics. GFP transcript levels relative to RPS17 were determined by RT-qPCR 48 h after transfection. One-way ANOVA with Tukey’s post-hoc multiple comparisons were used for data analysis. *p < 0.05; **p < 0.01. f The sequences of hsa-miR-21-5p and mutated residues (red residues) are shown below the graph. Error bars in all the graphs represent standard error of the mean (SEM) in three biological replicates.