(A–D) C57BL/6 and Mlkl^–/– mice were i.v. injected with ConA and analyzed 7 hours later. Experiments were repeated 3 times with similar results. (A) Plasma AST/ALT concentrations (n = 4 per group). ***P < 0.001 by paired Student’s t test. (B) Representative images of liver sections stained by TUNEL assay. (C) Quantification of necrotic areas in TUNEL assay–stained livers of ConA-challenged mice (n = 4 per group). ***P < 0.001 by paired Student’s t test. (D) Quantification of Ifng transcripts in liver lysates of mock- or ConA-challenged control and Mlkl^–/– mice by qPCR (n > 3 per group). ***P < 0.001 by paired Student’s t test. (E–I) 1 × 10⁷ bone marrow cells isolated from control or Mlkl^–/– mice were i.v. injected into lethally irradiated C57BL/6 recipient mice. Eight weeks later, mice were injected with ConA (n = 5 per group). (E) Plasma AST/ALT concentrations (n = 5 per group). (F) Representative images and quantification (n > 4 per group) of TUNEL assay–stained tissue sections. (G) Quantification of serum IFN-γ concentrations and hepatic Ifng transcripts in ConA-treated animals (n = 5 per group). (H) Quantification of hepatic Mlkl and Tnfa transcripts (n = 5 per group). (I) Quantification of TNF-α in supernatants of splenocytes isolated from ConA-challenged mice and stimulated ex situ with PMA/ionomycin for 24 hours (n = 5 per group). Error bars indicate +SD, gene expression levels are shown relative to Hprt. Scale bar: 250 μm. Original magnification in (B and F): ×7.55.