Fig. 5. The Loxl3-S704D mutant confers resistance to Oxaliplatin treatment in sleeping beauty transposon-induced liver cancer.

a, b For Loxl3-S704D mutant mouse generation, CRISPR–Cas9 mRNA, sgRNA and mutation donor ssDNA mixture were injected into the cytoplasm of fertilized eggs transferred into oviducts of pseudo pregnant female mice at 0.5 dpc for the birth of mutated mice. c 6-week-old male WT or Loxl3-S704D mutant mice were injected with sleeping beauty (SB) transposons carrying oncogenic dNβ-catenin and c-Met in the tail. The mice received low-dose Oxaliplatin (1 mg kg⁻¹ bodyweight) treatment intraperitoneally 3 times a week for 14 days. n = 4 in each group. d–j After 6 weeks induction and the following treatment, the representative livers of WT and Loxl3-S704D mutant mice are displayed, and liver weight was measured (d). HE staining was performed to ascertain tumor occurrence (e). The livers were further used for determinations of ALT activity (f), WB using indicated antibodies (g) and the lipid peroxidation levels (h). The ratio of tumor to liver mass (i) and the average tumor number (j) were analyzed and calculated. n = 4 in each group. k Another batch of mice receiving intraperitoneal low-dose Oxaliplatin (1 mg kg⁻¹ bodyweight) treatment was used to calculate the survival time by Kaplan–Meier plotter. n = 10 in each group. For (d, f, h–k), data represent means ± SEM of individual mouse groups (n = 4 or 10) and the statistical analysis was calculated by two tailed Student’s t test (d, i, j), one-way ANOVA with Tukey’s HSD post hoc test (f, h) or log-rank test (k). Source data are provided as a Source Data file. See also Supplementary Fig. 5.