Figure 2.

Neither ALT2 inhibition nor ALT2 knockout worsen liver injury after APAP overdose. (A–C) WT male mice 8–10 weeks old were treated with 100 mg/kg dexamethasone (Dex) followed by 300 mg/kg APAP 24 h later. Liver tissue was collected 6 h post-APAP. (A) ALT2 induction in liver homogenates was measured by immunoblot. (B) 5 μm-thick liver sections were glass-mounted and stained with hematoxylin and eosin (H&E) to measure necrosis. Representative images are shown at 100× magnification. (C) Necrosis was quantified in the liver sections. (D–H) WT male mice 8–10 weeks old were treated with 100 mg/kg BCLA followed by 300 mg/kg APAP 24 h later. Serum and liver tissue were collected 6 h post-APAP. ALT activity was measured in (D) serum and (E) liver tissue homogenates. (F) 5 μm-thick liver sections were glass-mounted and stained with H&E to measure necrosis. Representative images are shown at 100× magnification. (G) Necrosis was quantified in the liver sections. (I–M) Male liver-specific ALT2 KO mice and WT littermate controls 14–28 weeks old were treated with 300 mg/kg APAP or PBS vehicle. Serum and liver tissue were collected at 6 h post-APAP. (I) ALT2 was measured in liver homogenates by immunoblot. (J) Serum ALT was measured in APAP and PBS treated WT and KO mice. (K) Serum LDH was measured in APAP and PBS treated WT and KO mice. (L) 5 μm-thick liver sections were glass-mounted and stained with H&E to measure necrosis. Representative images are shown at 100× magnification. (M) Necrosis was quantified in the liver sections. Comparison of two means was performed by two-tailed t-test. Comparison of three or more groups was performed by ANOVA with post-hoc Tukey's test for multiple comparisons. ∗p < 0.05, ∗∗∗p < 0.001. In all panels, lines and error bars indicate mean ± SE. Circles show individual data points and sample sizes. Box plots show medians and IQR.