Research Paper Volume 14, Issue 13 pp 5449—5463
Long non-coding RNA H19 contributes to spinal cord ischemia/reperfusion injury through increasing neuronal pyroptosis by miR-181a-5p/HMGB1 axis
Figure 5. MiR-181a-5p targeted and negatively regulated HMGB1. (A) The sequences of HMGB1 3ʹUTR containing the miR-181a-5p binding sites or mutant binding sites were showed. (B) The expression of HMGB1 in the mouse spinal cord tissues following I/R. (C, D) Luciferase reporter gene assay was used to detect the luciferase activities of HMGB1-WT and HMGB1-MUT. (E) RIP assay was performed to determine the enrichment of miR-181a-5p and HMGB1 in Anti-Ago2 or IgG. (F) The correlations of the H19 and HMGB1 expression levels in the mouse spinal cord tissues following I/R were analyzed by Pearson correlation analysis. (G) The expression of HMGB1 was examined in PC12 cells transfected with miR-181a-5p mimic, miR-181a-5p inhibitor and their negative controls. (H, I) The expression of HMGB1 was examined in PC12 cells transfected with pcDNA-H19, sh-H19 and their negative controls. The results were presented as the mean ± SD. N = 3; *P<0.05.