Figure 4.

Sema4D is a direct target of Sja-miR-71a. (a). Three databases (miRanda, PITA, and TargetScan) were used to identify potential Sja-miR-71a targets. (b). mRNA expression of nine exemplary predicted targets of Sja-miR-71a in mice livers was analysed by qRT-PCR (n = 5–7 per group). (c). Sema4D in serum of schistosomiasis japonicum patients were analysed by qRT-PCR (n = 10–17 per group) and western blotting (n = 3–6 per group). (d). The wild-type m-Sema4D-3′- untranslated region (UTR) was cloned into psi-CHECK-2, and five predicted binding sites of Sja-miR-71a in the 3′-UTR of Sema4D gene; Dual-luciferase reporter assay performed on 293 T cells transfected with Sema4D UTR reporter plasmid together with Sja-miR-71a mimic or control mimic. (e). HSCs (LX2) were treated with S. japonicum egg-derived EVs (20 μg/mL) for 24 h. Sja-miR-71a and Sema4D expressions were analysed by qRT-PCR. (f, g). Expression of Sema4D in mice livers was analysed by immunohistochemistry (the sum of the IOD was analysed by Image-Pro Plus 6.0, n = 5–8 per group) and western blotting. Results are shown as mean ± SD. (b, e, f and g): One-way ANOVA Dunnett’s multiple comparison test. (c and d): Unpaired two-sample t-test.