Figure 6.

Necroptosis gene expression is up-regulated in the more differentiated epithelium of the ileum in the immature intestine. (A–D) Mouse intestinal epithelium was grown ex vivo as enteroids, which are maintained under either undifferentiated or differentiated conditions. (A and C) Bright-field and confocal micrographs of (A) undifferentiated or (C) differentiated enteroids in culture, stained with the indicated marker. Scale bar: 10 μm. (B and C) qRT-PCR of gene expression in (B) undifferentiated or (D) differentiated enteroids. Ascl2, Hopx, Lgr5, and Ki67 are markers of undifferentiated intestinal stem cells, whereas Sis, Muc2, ChgA, and Lyz1 are markers of differentiation as defined in the text. Undiff, enteroids grown in media to keep them stem-like (see the Materials and Methods section). Differentiation was for 72 hours. (E and F) qRT-PCR of necroptosis gene expression from enteroids (eg, epithelia) or intact bowel (ie, all cell types of that intestinal segment) in adult and juvenile animals. **P < .01 and ***P < .0001. Each dot is a separate sample. Representative of 3 separate experiments. DAPI, 4′,6-diamidino-2-phenylindole; Duo, duodenum; Ecad, E-cadherin; enteroids, juvenile epithelium; Ile, ileum; Jej, jejunum; RP, Rhodamine Phalloidin.