Extended Data Fig. 2: Death triggers induce varying degrees of mammalian cell death.

a, Cartoon schematic of in vitro cell line approach. b, Schematic of the CT26:FADD system. Doxycycline is used to induce expression of the construct, while addition of B/B induces oligomerization of the FKBP12 domains. Oligomerization leads to caspase activation and subsequent apoptosis. Cell death was measured using flow cytometry 5 h after B/B addition with or without QVD treatment. Membrane integrity was measured using DNA dye Sytox blue. n = 5 per condition. Western blot of the indicated apoptotic caspases (left), necroptotic machinery (right, top), or pyroptotic caspase 1 (right, below). DDR3 cells were used as positive controls for necroptosis. Representative blots from n = 3 independent experiments. c, CT26 cell death, as measured by flow cytometry. CT26:FADD cell death with or without caspase inhibition via QVD (left, n = 5 per condition) 5 h after death induction. CT26 cell death (centre, n = 4) 24 h after 600 mJ cm⁻² UV treatment. CT26 cell death (right, n = 3) 24 h after staurosporine treatment. CT26 cell death 24 h after 600 mJ cm⁻² UV exposure. The DNA dye 7AAD was used to assess membrane integrity. n = 4 per condition. CT26 cell death 24 h after 1 μM staurosporine treatment. The DNA dye 7-AAD was used to monitor membrane integrity. n = 3 per condition. Western blot of the indicated apoptotic caspases (left), pyroptotic caspase 1 (right, top), or necroptotic machinery (right, bottom). DDR3 cells were used as positive controls for necroptosis. Representative blots from n = 3 independent experiments. d, Bacterial CFU after anaerobic growth in UV irradiated CT26 supernatants (n = 4). Data are mean ± s.e.m. *P ≤ 0.05, **P ≤ 0.005, ***P ≤ 0.0005, ****P ≤ 0.0001, two-way ANOVA with Tukey’s multiple comparisons test (b), one-way ANOVA with Tukey’s multiple comparisons test (c, left) Student’s t-test (c, centre, right) or multiple two-tailed Student’s t-tests (d)

Source data.