Fig. 7

Lnc-LFAR1 promotes liver fibrosis through activating Notch signaling in primary HSCs. a The mRNA levels of Hes1, Notch2, Notch3, Jag1 and Hey2 were detected in lnc-LFAR1 downregulated primary HSCs by qRT-PCR. b Primary HSCs were infected with lentivirus-mediated shLFAR1 for 72 h and further treated with 10 ng ml⁻¹ TGFβ for additional 24 h. The protein levels of Nocth2, Notch3 and Hes1 were detected by western blot. GAPDH was used as an internal control. c, d The expression of Hes1, Notch2, Notch3, Jag1 and Hey2 was detected in lnc-LFAR1 over-expressed primary HSCs by qRT-PCR (c) and western blot d. GAPDH was used as an internal control. e, f Mice were treated with oil in combination with injection of lenti-NC (NC, n = 10), or CCl₄ in combination with injection of lenti-NC (NC+CCl₄, n = 10), or oil in combination with injection of lenti-shLFAR1 (shLFAR1, n = 10), or CCl₄ in combination with injection of lenti-shLFAR1 (shLFAR1+CCl₄, n = 10). The mRNA and protein levels of Hes1, Notch2, Notch3 and Hey2 were determined in liver tissues by qRT-PCR e and western blot f. GAPDH was used as an internal control. g Notch3 and Hes1 levels were detected by IHC. Scale bars, 400 μm for IHC (objective, ×10); 100 μm for IHC (objective, ×40). Right, five images of each liver and five livers from different mice were quantified for each group. Uncropped blots of this figure accompanied by the location of molecular weight markers are shown in Supplementary Fig. 18. In a, c and e, the number of biological replicates for each experiment was n ⩾ 3. Data are presented as means ± s.e.m. P values were analyzed by Student’s t-test in a and c, and by one-way analysis of variance followed by post hoc comparison in e and g. */#P < 0.05. *P < 0.05 vs shRNA-control in a; and *P < 0.05 vs LV-Control in c; and *P < 0.05 vs NC, #P < 0.05 vs NC+CCl₄ in e and g