Generation of Mlkl-deficient mice. (A) Targeting strategy for mutation introduction in the exon 1 of Mlkl locus by TALEN. The DNA-binding sites of the TALENs are indicated in red. (B) Summary of the generation of Mlkl-deficient mice. (C) Sequencing chromatograms showing nucleotide mutation introduced in the Mlkl locus adjacent to the FOKI cleavage site. Arrow indicates a double peak. (D) Western blot analysis of Mlkl protein levels in MEFs, BMDMs and peritoneal macrophages (MΦ) obtained from wild-type, Mlkl+/− and Mlkl−/− mice. (E) Western blot analysis of Mlkl protein expression levels in different organs of the wild-type and Mlkl−/− mice.