Skip to main content
Fig. 4 | BMC Cancer

Fig. 4

From: Mmu-miR-125b overexpression suppresses NO production in activated macrophages by targeting eEF2K and CCNA2

Fig. 4

Validation of mmu-miR-125b targets. a-b Alignment of potential mmu-miR-125b binding sites and mutations in the 3’ UTR of CCNA2 and eEF2K mRNA in mus musculus. c-d The intact or mutant 3’ UTR of the indicated genes were cloned into the psiCHECK2.2 luciferase reporter vector and then co-transfected with a mmu-miR-125b expression vector (miR-125b) or pLL3.7 (control) into 293T cells. Luciferase activity was analyzed 48 h after transfection using a dual luciferase reporter assay. 125b positive means the psiCHECK2.2 luciferase reporter vector include a sequence totally combined to miR-125b seed sequence. The results are expressed as the relative luciferase activity (firefly/renilla luciferase). The data are presented as the mean ± SD (n = 3) of three independent experiments. e The protein levels of CCNA2 and eEF2K in 24 h after 1 μg/ml LPS-activated RAW264.7-miR-125b and control cells were determined by western blot; GAPDH served as the loading control. f The protein levels of CCNA2 and eEF2K in 1 μg/ml LPS-activated RAW264.7 cells at different time points were determined by western blot; GAPDH served as the loading control. **p < 0.01; *p < 0.05

Back to article page