Fig. 6: Prolonged Gclc deletion in the liver induces a sex-dependent requirement of NRF2.

A Schematic for extended monitoring of Gclc deletion (Gclc L-KO), Nrf2 deletion (Nrf2 L-KO), and Gclc-Nrf2 deletion (L-DKO) in mice. Liver-specific gene knockout was induced with AAV-TBG-Cre. Mice were sacrificed 10 weeks post AAV-TBG-Cre injection unless humane endpoints were reached earlier. B Percent survival for WT (n = 8), Gclc L-KO (n = 6), Nrf2 L-KO (n = 6) and L-DKO (n = 12) mice following treatment with AAV-TBG-Cre. C Percent survival for female (n = 6) and male (n = 7) L-DKO mice following treatment with AAV-TBG-Cre. D Hepatocellular Injury Score from H&E-stained slides of the liver from female WT (n = 5), Gclc L-KO (n = 8), Nrf2 L-KO (n = 3), L-DKO (n = 4) and male WT (n = 5), Gclc L-KO (n = 8), Nrf2 L-KO (n = 3), L-DKO (n = 7) mice. A two-way ANOVA with subsequent Tukey’s multiple comparisons test was used to determine statistical significance (Female: WT vs. Gclc L-KO P value = 0.2504, WT vs. Nrf2 L-KO P value = 0.9999, WT vs. L-DKO P value = 0.5546, Gclc L-KO vs. L-DKO P value > 0.9999; Male: WT vs. Gclc L-KO P value = 0.1238, WT vs. Nrf2 L-KO P value > 0.9999, WT vs. L-DKO P value < 0.0001, Gclc L-KO vs. L-DKO P value = 0.0006; Female L-DKO vs. Male L-DKO P value = 0.0020). E Representative H&E-stained slides of the liver from female and male WT, Gclc L-KO, Nrf2 L-KO, and L-DKO mice 10 weeks following treatment with AAV-TBG-Cre. Scale bars = 200 µm. Data shown are representative of at least 3 replicates. F Serum triglyceride concentration of WT (n = 14), Gclc L-KO (n = 19), Nrf2 L-KO (n = 5) and L-DKO (n = 17) mice 10 weeks following treatment with AAV-TBG-Cre. A one-way ANOVA with subsequent Tukey’s multiple comparisons test was used to determine statistical significance (WT vs. Gclc L-KO P value < 0.0001, WT vs. Nrf2 L-KO P value = 0.7838, WT vs. L-DKO P value < 0.0001, Gclc L-KO vs. L-DKO P value = 0.1525). G Epididymal fat adipose tissue (eWAT) mass normalized to body mass from WT (n = 10), Gclc L-KO (n = 16), Nrf2 L-KO (n = 6) and L-DKO (n = 17) mice 10 weeks post-treatment with AAV-TBG-Cre. A one-way ANOVA with subsequent Tukey’s multiple comparisons test was used to determine statistical significance (WT vs. Gclc L-KO P value = 0.0112, WT vs. Nrf2 L-KO P value = 0.9675, WT vs. L-DKO P value = 0.0515, Gclc L-KO vs. L-DKO P value = 0.8936). Indicated n values represent biologically independent samples from mice. Data are shown as mean ± SEM. ns = not significant, * P value < 0.05, ** P value < 0.01, *** P value < 0.001, **** P value < 0.0001. (A) created with BioRender.com, released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.