Fig. 4: EFHD2 protects intestinal epithelium from TNF-induced cell death.
a Representative images of immunofluorescence for cleaved caspase-3 (green) and quantification of the small intestine from Efhd2f/f and Efhd2f/f Vil1cre/+ mice 4 h after intravenous injection with TNF. Left scale bar, 1000 μm; right scale bar, 100 μm. n = 6 each group. b H&E staining of the small intestine from a and quantification of villus height. Left scale bar, 100 μm; right scale bar, 200μm. n = 6 each group. c Time-series images of WT and Efhd2-/- enteroids on day 5 treated with 20 ng/ml TNF in the presence of a caspase-3/7 activity detection reagent (green). Digits represent hours: minutes; scale bars, 50 μm. Quantification of caspase-3/7 intensity was shown in the right histogram. AU arbitrary units. n = 15 enteroids each group. d Flow cytometric analysis of caspase-3/7 activity from dispersed WT and Efhd2-/- enteroids treated with TNF for 24 hours. n = 3 mice each group. e Representative images of WT and Efhd2-/- enteroids on day 5 treated with 20 ng/ml TNF together with GSK’872 or with Z-IETD-FMK for 24 hours in the presence of propidium iodide (PI) (red). Scale bar, 50 μm. The PI intensity was quantified at the endpoint (below). n = 12 enteroids each group. f, g Relative percentage change of body weight (f), DAI score (g) of Efhd2f/f and Efhd2f/f Vil1cre/+ mice subjected to 2.5% DSS and injected intraperitoneally with control IgG or anti-TNF antibody. h Representative images of colon morphology and lengths from mice in f on day 7. i, j Representative images of H&E staining (i) and histologic scores (j) of distal colons of mice in f on day 7. Upper scale bar, 100 μm; lower scale bar, 200 μm. n = 6 per group (f–h, j). Data are representative of at least three independent experiments; error bars show means ± s.d. P values were determined by unpaired two-tailed t-test. For body weight curves and DAI score, two-way ANOVA analysis with Sidak’s multiple comparisons test. Source data are provided as a Source Data file.
