miR-365 promotes HOXA9-mediated differentiation of mesenchymal stem cells to nucleus pulposus cells by interacting with HIF-1α

Figure 3 miR-365 binds 3’-UTR of Hoxa9, contributing to increased level of HIF-1α. A: Luciferase activity assay in mesenchymal stem cells (MSCs) expressing HOXA9-3’UTR-WT with miR-365 mimic or Hoxa9-3’UTR-Mut with miR-365 mimic (n = 4 per group); B and C: Relative mRNA level of Hoxa9 (B) and Hif-1α (C) in MSCs transfected with miR-365 mimic or antisense inhibitor (n = 3 per group); D: Relative protein level of HOXA9 and HIF-1α in MSCs transfected with miR-365 mimic or antisense inhibitor; E and F: Relative mRNA level of Hoxa9 (E) and Hif-1α (F) in MSCs transfected with Hoxa9 plasmid or siRNA (n = 3 per group); G: Protein level of HOXA9 and HIF-1α in MSCs transfected with Hoxa9 plasmid or siRNA; H: Quantitative PCR analyses of chromatin immunoprecipitation assay of HOXA9 binding to the Hif-1α promoter in MSCs (n = 3 per group). Data are presented as the mean ± SD. ^aP < 0.001, ^bP < 0.01, and ^cP < 0.05.