Fig. 5: EFHD2 suppresses RIPK1-dependent and -independent TNF-induced intestinal cell apoptosis. | Nature Communications

Fig. 5: EFHD2 suppresses RIPK1-dependent and -independent TNF-induced intestinal cell apoptosis.

From: EFHD2 suppresses intestinal inflammation by blocking intestinal epithelial cell TNFR1 internalization and cell death

Fig. 5

a Western blot analysis of lysates from wild-type and EFHD2 interfered HCT-116 cells treated with 20 ng/ml TNF alone, TNF and Cycloheximide (TC) or TNF and LCL-161 (TS) for the indicated time. Corresponding quantifications of cleaved-caspase-8, cleaved-caspase-7, and cleaved-caspase-3 by ImageJ were shown below. n = 3. Representative flow cytometric analysis of cell apoptosis (b) and caspase-3/7 activity (c) from WT and EFHD2-/- HT-29 cells treated with TC or TS for the indicated time and quantification of proportions of annexin V+ PI- apoptotic cells and the percentage of active caspase-3/7 cells (d). n = 3. e Western blot analysis of lysates from WT and EFHD2-/- HT-29 cells treated with TS and zVAD-fmk for indicated time. Quantifications of p-RIPK1, p-MLKL, and cleaved-caspase-7 were shown below. n = 3. f Western blot analysis and quantifications of p-IKKα/β and p-P65 in lysates from WT and EFHD2-/- HCT-116 cells treated with TNF for the indicated time. n = 3. Data are representative of three independent experiments; error bars show means ± s.d. P values were determined by unpaired two-tailed t-test in d. Two-way ANOVA analysis with Sidak’s multiple comparisons test in a, e, f. Source data are provided as a Source Data file.

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