(A) Percentages of apoptotic cells after uc.230 silencing. Caco-2 cells were transfected with the anti-uc.230 or control oligo (control); 48 hours after transfection, apoptosis was measured following 4 hours of treatment with TNFα (20 ng/mL) plus CHX (10 μg/mL). Values are means ± SEM (n = 4 or 5). *P < 0.05 compared with controls. (B) Images of TUNEL staining in cells described in A. Experiments were performed 3 separate times and showed similar results. Original magnification, ×200. (C) Changes in levels of caspase-3 in cells described in A. Whole-cell lysates were harvested, and the levels of total and cleaved procaspase-3 were assessed by Western blot analysis. GAPDH immunoblotting was performed as an internal control for equal loading. (D) Cleaved caspase-3 staining in enteroids after uc.230 silencing ex vivo. Enteroids were derived from the small intestines of WT mice and transfected with anti-uc.230 or control oligo (control). Enteroids were exposed to TNFα (10 ng/mL) plus CHX (5 μg/mL) 48 hours after transfection, and apoptotic cell death was examined 3 hours thereafter. Scale bars: 100 μm. (E) Quantitative data of cleaved caspase-3⁺ cells in enteroids described in D. Values are means ± SEM (n = 4 or 5). *P < 0.05 compared with controls. (F) Changes in levels of cleaved caspase-3 in enteroids described in D. (G and H) Apoptotic cell death in enteroids exposed to LPS after uc.230 silencing. Enteroids were treated with LPS (50 μg/mL) and changes in the levels of cleaved caspase-3 were examined 24 hours thereafter. *P < 0.05 compared with controls (n = 5). Scale bars:100 µm. In A, E, and H, statistical significance was analyzed using unpaired, 2-tailed Student’s t tests.