Figure 3.

miR-27a-3p Targets the 3′ UTR of ATF3

(A) Atf3 transcript level in primarily cultured rVSMCs treated with Pi. The results were obtained from an mRNA microarray (GEO: GSE74755). (B) miR-27a-3p mimic significantly attenuated the luciferase activity driven by Atf3-3′ UTR. psiCHECK2-Atf3-3′ UTR was used for luciferase activity measurement. (C) miR-27a-3p inhibitor dramatically increased the luciferase activity. (D) miR-27a-3p mimic failed to inhibit mutant Atf3-3′ UTR where the miR-27a-3p-binding sequence was altered. (E) miR-27a-3p mimic significantly reduced the mRNA transcript amount of Atf3. (F and G) miR-27a-3p mimic reduced the protein amount of Atf3 as determined by western blot analysis. Quantification results from the western blot (F) are shown in (G). Error bars indicate SD. ∗p < 0.05, ∗∗p < 0.01. NS, not significant.