Figure 3.

Knockdown of Fendrr attenuates CCl₄-induced liver fibrosis. Mice were injected with oil (oil, n = 5), CCl₄ (CCl4, n = 10), CCl₄ in combination with injection of control lentivirus (CCl4+shNC, n = 10), and CCl₄ in combination with injection of lentivirus capable of expressing Fendrr shRNA (CCl4+shFendrr, n = 10). The lentivirus was injected once at 2 days before the first CCl₄ injection. A, liver fibrosis was evaluated by H&E staining, sirius red staining as well as collagen I immunohistochemical staining and α-SMA immunofluorescence staining. The scale bar represents 100 μm. B, quantification of the sirius red positive area. C, quantitative evaluation of hepatic hydroxyproline. The hydroxyproline contents are expressed as μg/mg wet liver weight. D, Fendrr expression was examined by qRT-PCR. E, quantitative evaluation of Col I staining area. F, quantification of α-SMA staining area. G, hepatic collagen I and α-SMA were detected by Western blot (left). Relative protein level was calculated, respectively, by band intensity against β-tubulin (right). H, hepatic Tgfb1, Pdgfa, Col1a1, and Timp1 mRNA expressions were examined by qRT-PCR. The results are shown as fold change compared with oil group mice. I, knockdown of Fendrr suppresses the increase of serum ALT levels in CCl₄-treated mice. Data are the mean ± SD of at least three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. α-SMA, smooth muscle α-actin; ALT, alanine aminotransferase; Col I, collagen I; qRT-PCR, quantitative real-time PCR; TIMP, tissue inhibitors of metalloproteinase.