Fig. 3

Functional assay. a Gain-of-function assay using an exogenous mimic-miR-21 was used to perform an initial screen of miR-21 target genes. q-PCR analyses of KRIT1, FOXO1, NEFL2L, SOD2, SOD1, CAT and GPx-1 mRNA relative expression levels were performed in HUVECs cultured in normal glucose (5 mmol/L) for 24 h and compared to scrambled control. A two-tailed t-test was applied for statistical analysis. *p < 0.05, ***p < 0.001, both vs scrambled control. b Luciferase assays tested the binding between 3′-UTR SOD2 and miR-21. The luciferase data are expressed as a ratio between firefly and renilla luciferase to normalize for the transfection variability between each sample and subsequently normalized against lysate protein levels to account for differences in the total protein concentration. Measurements were carried out in quadruplicate for three independent experiments. **p < 0.01, ANOVA 1-way, followed by Tukey’s multiple comparison test