Allogeneic T cells induce TNF-α–mediated necroptosis in intestinal organoids. (A) Fold change in indicated cytokines in culture supernatants from Figure 3B. Each value is normalized to nonstimulated samples; n = 3 mice each. (B) Viability of small intestinal organoids treated or not with anti–TNF-α and/or anti–IFN-γ antibody and cocultured with B10.BR T cells for 48 hours; n = 3 mice each. Representative images of cocultured small intestinal (C) and colonic (E) organoids. Arrowheads denote dead organoids. Scale bars, 100 µm. Viability of small intestinal (D) and colonic (F) organoids from B6-background Atg16L1^f/f (f/f), Atg16L1^ΔIEC (ΔIEC), f/f Ripk3^−/−, and ΔIEC Ripk3^−/− mice cocultured for 48 hours with B10.BR T cells; n = 3 mice each. Data points in A, B, D, and F are mean of technical replicates. Bars represent mean ± standard error of the mean, and ≥2 independent experiments were performed. *P < .05, **P < .01, ***P < .001, ****P < .0001.