HK reconstitution exacerbates liver injury. (A) Western blot of plasma HK in deficient (HK−/−), reconstituted (HK−/− + hHK), and sufficient (HK+/+) mice. Green = polyclonal antibody against the light chain of hHK. Red = polyclonal antibody against the FXI- and PK-binding domain within the light chain of mouse HK. Arrow indicates full-length HK (∼120 kDa). (B) Circulating levels of ALT in plasma from HK-deficient animals, HK-deficient animals reconstituted with hHK, and HK-sufficient animals. (C) Injury score calculated based on criteria in supplemental Table 1, three 100× fields per mouse. (D) Representative photomicrographs indicating necrotic area in HK−/−, HK−/− + hHK, and HK−/− mice. (E-H) Quantitation of plasma concentration of cytokines as indicated. N = 6 mice per group. All data collected at 24 hours after APAP administration. Error bars represent standard deviation of the mean. *P < .05, **P < .01, ***P < .001, ****P < .0001. G-CSF, granulocyte colony-stimulating factor; MIP-1β, macrophage inflammatory protein-1β.