Figure 5. H19 upregulates HPRT1 expression by inhibiting miR-301b-3p. (A) The predicted miRNAs binding to H19 and HPRT1; (B) Putative binding sites between miR-301b-3p and HPRT1 3’UTR; (C) Putative binding sites between miR-301b-3p and H19; (D) The expression of miR-301b-3p and H19 in N27 dopaminergic neurons and substantia nigra tissues assessed by RT-qPCR. (E) Binding of miR-301b-3p and HPRT1 verified by dual-luciferase reporter gene assay. (F) N27 dopaminergic neurons transfected with miR-301b-3p-Bio or miR-301b-3p-Mut-Bio or NC-Bio tested by RNA pull-down assay 48 h after transfection. H19 expression was evaluated by RT-qPCR. (G) Verification of RIP assay for direct interaction of H19 with Ago2 protein. (H) The expression of H19 and miR-301b-3p in the substantia nigra tissues in response to oe-H19 examined by RT-qPCR. (I) The miR-301b-3p expression and the mRNA expression of HPRT1 in the substantia nigra tissues in response to oe-H19 in the presence of agomir-miR-301b-3p examined by RT-qPCR. *p < 0.05. Measurement data are by means ± standard deviation. Comparison between two groups was analyzed by unpaired t test, and comparison among multiple groups by one-way analysis of variance. n = 6.