Figure 6. MiR-181a increased the expression of fibrosis-related proteins and concentration of inflammatory cytokines through Egr1/TLR4 signaling. (A) Binding region between miR-181a and Egr1. (B) Luciferase reporter assay for the confirmation of direct binding relationship between miR-181a and Egr1 was performed with luciferase reporter plasmids of wild- or mutant-type Egr1; (C) Egr-1 mRNA expression was upregulated in MCs transfected with miR-181a inhibitor. (D) TLR4 mRNA expression was upregulated in MCs transfected with miR-181a inhibitor. (E) miR-181a inhibition increased the protein level of Egr-1 and TLR4. (F) The protein expression of α-SMA was measured in MCs by Western blot. Cells were transfected with inhibitor-miR-181a or pcDNA3.1-Egr1. (G) The concentration of IL-1β in MCs was measured by ELISA. The cells were transfected with miR-181a inhibitor or pcDNA3.1-Egr1.