Research Paper Volume 13, Issue 3 pp 4115—4137

Figure 3. XIST directly targets miR-199a-3p. (A) The putative binding sites of miR-199a-3p in XIST were predicted using DIANA tools. (B) HEK-293T cells were co-transfected with the miR-199a-3p mimics or inhibitor and XIST-WT or XIST-MUT luciferase reporter plasmids. Luciferase activities were measured 48 h after transfection. (C) The binding between miR-199a-3p and XIST was further confirmed by RIP analysis. (D) SH-SY5Y and PC-12 cells were transfected with shXIST. Transfection efficiency was assessed by qPCR analysis. (E) The effect of shXIST on miR-199a-3p expression was evaluated by qPCR. (F) XIST expression was inhibited by transfecting the SH-SY5Y and PC-12 cells with shXIST. (G) qPCR results showed that shXIST transfection improved miR-199a-3p expression, which was repressed by co-transfection with the miR-199a-3p inhibitor. The data are representative of three experiments. **p <0.01, ***p <0.001 and ns, not significant.