Fig. 6. Overexpression of circBNC2 inhibits liver fibrosis by attenuating hepatocytes arrested in G2/M phase.

a Mice were treated by tail vein injection of either AAV9-circBNC2 or AAV9-Vector 3 weeks before processing hepatic fibrosis model by injecting CCl₄ (once a week for 6 weeks). Mice were euthanized 6 weeks post the first injection of CCl₄. b, c Representative images of in situ hybridization of circBNC2 expression in livers from mice injected with AAV9-circBNC2 (b) and the quantification data (c). d, e Representative images of Immunohistochemistry of p-H3 in liver sections from mice treated with AAV9-circBNC2 and CCl₄ (d), and the quantification data (e). f, g Western blots showing expression of cyclin B1 and cyclin D1 in mice treated with AAV9-circBNC2 and CCl₄ (f), and the quantification data (g). h, i Representative images of TUNEL staining of apoptotic cells in the sections from CCl₄-treated mice overexpressed with circBNC2 (h), and the quantification data (i). j qRT-PCR showing the expression of Tgfb1 in livers from CCl₄-treated and circBNC2-overexpressed mice. k Western blots showing expression of αSMA and Col I in the liver homogenates from CCl₄-injected mice overexpressed with circBNC2. l–n Representative images of Immunohistochemistry of αSMA and Col I in liver sections from circBNC2-overexpressed mice treated with CCl₄ (l) and the semi-quantification data (m, n). o, p Representative images of Masson staining of liver sections from CCl₄-injected mice with circBNC2 overexpression (o), and the quantification data (p). q Reduced hydroxyproline (HYP) in liver tissues in CCl₄-injected mice with circBNC2 overexpression. r Overexpression of circBNC2 improved liver function in CCl₄-injected mice, evidenced by decrease in serum alanine aminotransferase (ALT). For c, e, i, j, m, n, p–r, n = 6 in each group. Data are expressed as means ± SD. One-way ANOVA with Bonferroni post hoc test was used for comparison among multiple groups (c, e, g, i, j, m, n, p, q, r). Source data are provided as a Source Data file.