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. 2019 Oct 10;9(2):295–312. doi: 10.1016/j.jcmgh.2019.10.002

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© 2020 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

TNF induces apoptosis in IKKβ(EE)-expressing intestinal enteroids. (A) WT and Ikkβ(EE)^IEC enteroids were photographed under brightfield 4 hours after TNF stimulation (40 ng/mL). Original magnification ×200. (B) Ikkβ(EE)^IEC and Ikkβ(EE)^IEC/Tnf^–/– enteroids were photographed under brightfield before and after incubation with TNF (40 ng/mL). Original magnification ×200. (C) WT and Ikkβ(EE)^IEC enteroids were stained with cC-8 antibody and visualized by confocal microscopy at the indicated times after TNF (40 ng/mL) addition. Magnification bars: 100 μm. (D) WT and Ikkβ(EE)^IEC enteroids were TUNEL stained and analyzed by confocal microscopy before and after incubation with TNF. Apoptotic cells are bright green. Magnification bars: 100 μm. (E) Lysates of WT and Ikkβ(EE)^IEC enteroids were IB analyzed at different times after TNF addition. (F) Lysates of Tnf^-/- and Ikkβ(EE)^IEC/Tnf^–/– enteroids incubated with or without TNF were analyzed for caspase activation by IB.