Mice were treated with 300 mg/kg acetaminophen (APAP). After 24 hours, Ly6Chi F4/80neg monocytes were isolated from the livers of mice by using immunomagnetic beads. A and B: Nonparenchymal cells (A) and purified Ly6Chi F4/80neg cells (B) were analyzed by flow cytometry. C and D: Immunofluorescence was used to detect Ly6C (green; C) and F4/80 (red; D) in cultured Ly6Chi F4/80neg monocytes. E–G: Ly6Chi F4/80neg cells were treated with control media or media containing necrotic hepatocytes. Data are expressed as means ± SEM (E–G). n = 3 (E–G). *P < 0.05 versus control. Original magnification, ×400 (C and D). Ccl, chemokine (C-C motif) ligand; iNOS, inducible nitric oxide synthase; TNF-α, tumor necrosis factor-α.