Fig. 2.

NS-KO improves liver necrosis, regeneration, and survival in vivo. a Generation of NS-KO mice. Left panel showing a schematic representation of NS-KO mice, NSun2 genomic locus, and NSun2 deleted locus. The targeting sgRNA was designed to knock out 56 bp DNA fragments in exon 2 of the NSun2 gene. The right panel is genotyping analysis of WT and NS-KO mice confirmed by western blotting. b H&E staining showing mouse liver in WT and NS-KO mice not subjected to stress. Scale bar, 50 μm; n = 3 mice. c Schematic of the construction of liver injury mice models. Liver injury was induced by repeated injections of CCl₄. The part above the arrow line of time represents the injection time, and the part under the arrow line represents the liver-obtained time. D0: no injury; D3: day 3 after injury; D27: day 27 after injury. d H&E staining showing hepatic necrosis at different time points after CCl₄ injection. Dotted lines indicate the necrotic or inflammatory area and the percentage of the necrotic or inflammatory areas is quantified. Scale bar, 50 μm; n = 3 mice. e, f Mouse serum ALT (e) and AST (f) concentrations were assessed to determine the degree of liver injury after repeated CCl₄ injection, n = 4 mice. g Hepatocytes proliferation in liver sections. Three hours before the mice were euthanized, 5-bromodeoxyuridine (BrdU) incorporation was performed for WT and NS-KO mice treated with CCl₄ and BrdU-positive hepatocytes were quantified. Scale bars, 200 μm; n = 3 mice. h TdT-mediated dUTP nick end labeling (TUNEL) assay on liver sections of WT mice or NS-KO mice treated with CCl₄. Green cells indicate TUNEL-positive (apoptotic) cells. Nuclei were stained with DAPI, and TUNEL-positive nuclei were quantified. Scale bars, 200 μm; n = 3 mice. i Proliferation- and survival-related gene expression was quantified by qRT-PCR after liver injury. n = 3 mice. For data in this figure, representative images are shown. All data are presented as the mean ± SD. *P < 0.05, **P < 0.01, Student’s t-test