Figure 7: miR-294 and miR-721 bind to Nos2 3′UTR.

RAW 264.7 (5 × 105) were transfected with 5 μg of pmiRGLO, empty or with the regions containing miR-294 or miR-721 binding sites to Nos2 3′UTR, plus 100 nM of miR-294 or/and miR-721 mimics (A). After 24 h, the cells lysates were analyzed for luciferase activity (B). The values of firefly luciferase (FL) were firstly normalized by renilla luciferase (RL), and then normalized by the values obtained with cells transfected with pmiRGLO empty construct. Each bar represents the average ± SEM of the values obtained in 3 independent experiments (n = 5). Statistical significance was determined based on two-tailed Student’s t test. *p < 0.05, compared to pmiRGLO empty vector.