Figure 7. miR-223 regulates Cxcl10 and Taz expression in hepatocytes.

(A) Bioinformatics approach analyses of the target prediction of miR-223. (B) Dual-Luciferase activity assay was performed to verify binding between miR-223 and CXCL10 or TAZ. AML12 cells were co-transfected with control luciferase vector, Cxcl10 3’UTR vector or Taz 3’UTR vector and miR-223 mimics or non-specific (NS)-miRNA mimics for 48h. Relative luciferase activity was determined. (C) AML12 cells were transfected with NS-miRNA mimics and miR-223 mimics for 24h, and then challenged with 0.1 mM palmitic acid (PA) for 3h. The Cxcl10 and Taz mRNA levels were measured by RT-qPCR. (D) Primary hepatocytes from WT and miR-223KO mice were treated with PA for 3h. The Cxcl10 mRNA expression was measured by RT-qPCR. (E) AML12 cells were transfected with NS-miRNA mimics and miR-223 mimics for 24h. The YAP and TAZ proteins were measured by western blotting. (F) AML12 cells were transfected with NS-miRNA mimics and miR-223 mimics for 24h, and then treated with 0.1 mM PA for 24h. The protein levels in whole hepatocyte lysates were determined by Western blot analyses. Values represent means ± SEM from three independent experiments. *P< 0.05 as indicated; ^#P<0.05, ^###P<0.001 in comparison with control NS-miRNA mimic groups in panel C or 0h WT group in panel D.