Leukocytic infiltration (indicated by green arrows, B-C), lipogenic accumulation (indicated by stars *, B, H, K), nitroblue tetrazolium (NBT) deposits (blue precipitate indicated by green arrows, E), hepatic Kupffer cell (ED1) stain (brown stain, D-F), 4-hydroxy-2-nonenal (4-HNE) stain (brown color, H), and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) stain were used to demonstrate de novo production of oxidative stress in the D-GalN treated liver. Oxidative stress indicated by NBT (green arrows) appeared in the Kupffer cells (identified by ED1 brown color) and hepatocytes of rat livers subjected to 24 hr of D-GalN treatment (E), but less evident in the control liver (D) and the liver with low dose of GT pretreatment (F). 4-HNE stain for oxidized protein concomitantly occurred in the D-GalN treated hepatocyte with lipogenic accumulation (H), but not appeared in the control (G) or GT pretreated livers (I). TUNEL stain for apoptotic cell death was absent in the control liver (J) and GT treated liver (L), but appeared in the liver after 24 hr of D-GalN treatment (K). A-C, hematoxylin and eosin stain, × 400; D-F, NBT+ED1 counterstain, × 400; G-I, 4-HNE stain, × 600;J-L, TUNEL stain, × 400.