CCl4-induced liver fibrosis model

Male C57BL/6J mice aged eight weeks received an intraperitoneal injection of 10% CCl₄ in olive oil (7 μL/g/mouse) biweekly for six weeks. Similarly, the control mice were treated with olive oil by using the same method.

For the CCl₄-induced mouse liver fibrosis model, all C57BL/6J mice were randomly divided into three groups: mice treated with CCl₄ alone (n = 6), CCl₄ in combination with an injection of pNL-EGFP/CMV/WPREU3-NC (CCl₄+NC; n = 6), and CCl₄ in combination with an injection of pNL-EGFP/CMV/WPREU3-hsa_circ_0070963 (CCl₄+CircRNA; n = 6). Every two weeks, the lentivirus (1 × 10⁹ pfu/100 μL) was injected into the mice via the tail vein for a total of six weeks. All animals were provided by the Experimental Animal Center of Wenzhou Medical University. The animal experimental protocol was approved by the University Animal Care and Use Committee in The First Affiliated Hospital of Nanjing Medical University. At the end of the experiment, mice were sacrificed under anesthesia and the livers were obtained for further studies such as Masson staining and immunohistochemistry.