Fig. 4

ISM1 regulates necroptosis in the lungs. (A-D) Representative Western blot analysis of (A) pMLKL, (B) RIP3, (C) cleaved caspase 8, and (D) cleaved caspase 3 protein expression in lung tissues comparing WT (n = 5) and ISM1^−/− (n = 7) mice with and without HDM sensitization. Data are presented as the mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001, using one-way ANOVA. (E) Release of pan-necrotic marker LDH in the BALF (n = 4). (F) Representative photomicrograph of immunohistochemical staining of lung tissue sections against pMLKL. Bars = 50 μm. (G) Immunofluorescence staining of lung sections from HDM-challenged ISM1^−/− mice. Bars = 20 μm. (White arrows) Indicates colocalization between pMLKL and the AM marker CD68. (H) rISM1 regulates pMLKL expression in MH-S cells. Data are presented as the mean ± SD. *P < 0.05; **P < 0.01; ****P < 0.0001, using one-way ANOVA. pMLKL, phosphorylated mixed lineage kinase domain-like protein; RIP, receptor-interacting protein 3; LDH, lactate dehydrogenase, Nec1, necrostatin 1