Research Paper Volume 11, Issue 11 pp 3585—3600

Blocking lncRNA H19-miR-19a-Id2 axis attenuates hypoxia/ischemia induced neuronal injury

Figure 4. Direct interaction between H19 and miR-19a. (A) qRT-PCR analysis revealed that miR-19a levels in penumbra cortex of rats increased gradually and significantly within 72 h after MCAO/R. (B) Similar results were observed in OGD models. (C) With knockdown of H19 by siRNA, miR-19a level was significantly elevated in OGD neuronal cells. (D) The predicted fragment including miR-19a binding sites on H19 was cloned into a pmirGLO vector (H19-wt), and a mutated vector (H19-mut) was also generated by replacing the binding sites with its complimentary sequence. (E) Dual-luciferase report assay revealed that miR-19a mimic reduced the luciferase activity of H19-wt, but not of H19-mut. (F) RIP indicated that H19 was preferentially enriched in Ago2-containing bead compared to those harboring control immunoglobulin G (IgG) antibody. Furthermore, H19 was pulled down by biotin-labeled miR-19a oligos, but not the mutated oligos.