Fig. 6
lnc-Helf promotes hepatic inflammation and fibrosis through activating AKT pathway. A Mice were divided into six groups: AAV8-NC, NC + CCl4, lnc-Helf-sh1#, lnc-Helf-sh1# + CCl4, lnc-Helf-sh2#, and lnc-Helf-sh2# + CCl4. Western blot was used to determine the protein level of phos-AKT (Thr308), phos-c-Raf1 (Ser259), phos-GSK-3β (Ser9), and AKT in liver tissues of each group. B, C Primary HSCs at day 2 were infected with lenti-control, lnc-Helf-shRNA, or lenti-lnc-Helf for 48 h, following treatment with 10 ng/ml TGF-β1 for 24 h. Western blot was used to determine the protein level of phos-AKT (Thr308) and AKT. GAPDH was used as an internal control. D Primary HSCs treated with or without the AKT inhibitor MK2206 at day 2 were infected with lenti-control or lenti-lnc-Helf for 72 h. qRT–PCR was used to detect the RNA level of lnc-Helf, Acta2, Col1α2, Timp1, Pcna, and Cyclin E1. E, F RAW264.7 cells treated with or without the AKT inhibitor MK2206 were transfected with pcDNA3.1-lnc-Helf or pcDNA3.1 for 72 h. Mature supernatant IL-1β level was detected by ELISA (E); cell proliferation was detected by CCK8 (F). Data are presented as mean ± SEM. */#p < 0.05. *p < 0.05 for LV-control + DMSO or pcDNA3.1 + DMSO. #p < 0.05 for LV-lnc-Helf + DMSO or pcDNA3.1-lnc-Helf + DMSO, one-way ANOVA (D–F)