Extended Data Fig. 7: ASM cleavage is requires for IEC extrusion.
From: Caspase-7 activates ASM to repair gasdermin and perforin pores
a, Strategy to generate ASM D249A mutant mice by CRISPR–Cas9. Target sequence for guide RNA in exon 2 is shown in red. Repair oligo DNAs (200 nt) containing indicated mutation are also used for electroporation with Cas9. Successful mutation was confirmed by Sanger sequencing. b, Quantification of EpCAM+ cells per extruding site in non-infected WT, Smpd1DA/DA, and Casp7–/– caeca (n = 4 in each group). c–f, Live imaging of indicated IEC organoids, which were quantified for rupture percentage after PBS (c) or FlaTox (d) treatment, extrusion starting time after FlaTox treatment (e), and PI Intensity after TNF+CHX or PBS treatment (f). Data are pooled from 2 (b) or 3 (c-d) experiments or are representative of 3 experiments (f). For (e), WT (n = 27) and Smpd1DA/DA (n = 41) organoids pooled from 3 experiments were analysed. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (Two-sided unpaired t-test in (d, e), two-way ANOVA with Sidak’s post-hoc test in (f)). Data are shown as mean ± SEM. Exact p values in Source Data EDF7
