Extended Data Fig. 6: ZBP1 contributes to the early postnatal lethality of Adar1^−/− Mavs^−/− mice and induces cell death in MAVS-deficient MEFs.

a, Numbers of offspring genotyped at birth from intercrossing Adar1^WT/−, Adar1^WT/− Zbp1^−/−, Adar1^WT/− Mavs^−/− or Adar1^WT/− Zbp1^−/− Mavs^−/− mice. b, Representative images of Adar1^WT/− Mavs^−/− Zbp1^−/− (n = 6) and Adar1^−/− Mavs^−/− Zbp1^−/− (n = 8) mice at about 15 weeks of age. c, Relative body weight compared to littermate controls in 10-week-old mice with the indicated genotypes. d, RBC counts, HGB and HCT levels in blood of 15–20 week-old mice with the indicated genotypes. In c, d, control mice include littermates with Adar1^WT/− or Adar1^WT/WT genotypes. Dots represent individual mice, bar graphs show mean ± s.e.m and P values were calculated by two-sided nonparametric Mann-Whitney test. e, Representative H&E-stained sections from the spleen, liver, lung, heart, small intestine and colon of 15–20 week-old Adar1^WT/WT Mavs^−/− Zbp1^−/− (n = 5) and Adar1^−/− Mavs^−/− Zbp1^−/− (n = 5) mice. Black arrows indicate dying epithelial cells in small intestine and colon sections. Scale bars, 200 μm. f, Immunoblot analysis of total lysates from murine embryonic fibroblasts (MEFs) of the indicated genotypes, stimulated with IFNγ (1,000 U ml⁻¹) for 24 h. One representative out of two independent experiments is shown. GAPDH was used as a loading control. g, h, j, Cell death measured by DRAQ7 uptake in MEFs of the indicated genotypes treated with IFNγ (1,000 U ml⁻¹) alone or with combinations of IFNγ (1,000 U ml⁻¹) (24-h pretreatment), cycloheximide (CHX) (1 μg ml⁻¹), QVD-OPh (QVD) (10 μM) and GSK’872 (3 μM). Graph shows mean values from technical triplicates (n = 3), from one representative out of three independent experiments. i, Immunoblot analysis of total lysates from MEFs of the indicated genotypes pre-stimulated with IFNγ (1,000 U ml⁻¹) for 24 h followed by treatment with CHX (1 μg ml⁻¹) for the indicated time points. Data are representative of two independent experiments. GAPDH was used as loading control. For gel source data, see Supplementary Figure 1.

Source data