Figure 4.

MiR-223 inhibits PDGF signaling pathway in HSCs by targeting Pdgfra and Pdgfrb. (A-C) WT and miR-223KO mice were treated with CCl₄ twice a week for 4 weeks or 8 weeks. (A) RT-qPCR analyses of Pdgfra and Pdgfrb. (B) Liver tissues were subjected to western blot analyses of PDGFRα and PDGFRβ. (C) Representative images of PDGFRα and PDGFRβ staining are shown. (D) Using a miRNA database (http://www.targetscan.org/), putative binding sites of miR-223 were predicted in the 3'-UTR of mouse Pdgfra and Pdbgrb mRNAs. (E) Primary mouse HSCs were transfected with miR-223 mimics and nonspecific miRNA mimics (NS), and the mRNA levels of Pdgfra and Pdgfrb were detected by RT-qPCR analyses. (F) Primary mouse HSCs were transfected with miR-223 mimics and nonspecific miRNA mimics (NS) for 24 hours followed by treatment with vehicle or PDGFBB (10 ng/ml) for 20 min. Total cell lysates were subjected to western blot analyses of PDGF signaling pathway. The blots were quantified (right panel). Values represent means ± SEM. In panels A-C, n=5-8/group; Panels E-F from three independent experiments. ^*P< 0.05, ^**P< 0.01, ^***P< 0.001.