Fig. 5.

OL upregulates miR-519d expression through the hypoxia response element (HRE) in its promoter region. a Promoter region of miR-519d contains putative hypoxia response element (HRE). Wild type (HRE-wt, GCGTG) or mutated (HRE-mut, ATACA) HRE sites from miR-519d promoter were cloned at the upstream of a luciferase reporter gene open reading frame (Luc ORF). b HNE-1 cells were incubated for 24 h with media containing 0 or 100 μM CoCl₂, respectively, followed by ChIP assay using control IgG or HIF1α antibody (anti-HIF1α). c Luciferase activities of HRE-wt or HRE-mut constructs were measured in HNE-1 cells treated as in (b). d HNE-1 cells were incubated for 24 h with media containing 0 or 200 μM of OL, respectively, followed by ChIP assay using control IgG or HIF1α antibody (anti-HIF1α). e Luciferase activities of HRE-wt or HRE-mut constructs were measured in HNE-1 cells treated as in (d). Values were expressed as mean ± SD. **p < 0.01, ***p < 0.001, ns1 not significant, compared to control. #p < 0.05, ##p < 0.01, ns2 not significant, compared to 0 μM OL