Fig. 3: Tipe0−/− enterocytes have dysregulated growth and responses to injury. | Nature Communications

Fig. 3: Tipe0−/− enterocytes have dysregulated growth and responses to injury.

From: TNFAIP8 controls murine intestinal stem cell homeostasis and regeneration by regulating microbiome-induced Akt signaling

Fig. 3: Tipe0−/− enterocytes have dysregulated growth and responses to injury.The alternative text for this image may have been generated using AI.

D7 enteroid survival (by CTG) after exposure to 24 h of 100 ng/mL TNF (a) or 24 h of hypoxia (b p = 0.0224); Total N = 13/group for a; total N = 18 for WT and 17 for Tipe0−/− for b; pooled from three independent experiments. c CTG survival assay 24 h after exchange into media lacking supplemental growth factors (N = 18/group, pooled from two independent experiments, p = 0.0205) with a 5-day survival curve by manual counting (N = 47 (WT), 23 (Tipe0−/−); ****p < 0.0001 by Mantel–Cox test), with images (d) of the time course and development of de novo Tipe0−/− spheroids 1 day after growth factor withdrawal (e, white spheres). Bars = 1090 µm, image representative of three independent experiments, insert 5× magnified. f Day 7 enteroid cultures morphology. Bars = 1110 µm. Image representative of 5 independent experiments. g D7 enteroid diameters; N = 29 for WT, 30 for Tipe0−/−. h Viable D7 enteroids after equal seeding, N = 3/group; p = 0.0048. i Viable ileal crypts; N = 9/group pooled across three independent experiments. j Ki-67 staining with quantification in k; staining from two independent experiments were pooled. Total N = 6 mice (WT), 5 (Tipe0−/−); p = 0.0087 Images representative of the mean staining in each group. Bars = 110 µm. Inserts are 3× magnified. l–o Brdu/Edu pulse chase; N = 3 mice for WT, 4 for Tipe0−/− and staining was repeated twice. Quantification in l: three fields of view/mouse, total N = 9 (WT), 12 (Tipe0−/−), p = 0.067 for Edu, p = 0.005 for Brdu. Images shown in m representative of staining in 3/4 mice in WT (Brdu failed to incorporate in one mouse), 4/4 mice in Tipe0−/−; bars = 130 µm. Distance migrated by Edu+ and Brdu+ cells (white brackets), with quantification in n; N > 60 measurements/group and three random images per section analyzed. Box & whisker plots range from minima to maxima, with median as the center, and the bounds of the box from 25–75% percentile. O) Crypt-villi length from N = 19/group pooled from 4 mice/group. For all graphs, error bars are mean ± SEM, *p < 0.05; **p < 0.01, ***p < 0.001; ****p < 0.0001. Analyses done by two-tailed Mann–Whitney U test unless otherwise indicated. Source data are provided as a source data file.

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