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. 2011 May 9;108(21):8743–8748. doi: 10.1073/pnas.1019574108

Fig. 3.

Fig. 3.

Epithelial cell MyD88 is required for RegIIIγ expression in small-intestinal γδ IEL. RegIIIγ was quantified by (A) Q-PCR of sorted small-intestinal γδ IEL and (B) flow cytometry of total IEL, performed as in Fig. 1. Gated γδ IEL populations are shown, and percentages of gated populations are given. Four to five mice were pooled per group; results are representative of three independent experiments. (C) Schematic of bone marrow chimera experiment. Ly5.1 wild-type mice were irradiated and reconstituted with bone marrow cells from Ly5.2 MyD88−/− mice. The chimeric intestines retained residual recipient cells while also supporting engraftment of transplanted cells (40% donor/60% recipient γδ IEL). EC, epithelial cells. (D) Eight weeks after reconstitution, Ly5.1 and Ly5.2 γδ IEL were isolated by FACS and analyzed individually for RegIIIγ mRNA by Q-PCR. Each point represents an individual mouse, and the conventional experimental groups were cohoused for 5 d before sacrifice to ensure a shared microbiota. (E) Q-PCR for RegIIIγ expression in sorted small-intestinal γδ IEL. MyD88ΔIEC, mice with an epithelial cell-specific MyD88 deletion; MyD88fl/fl, littermates harboring two floxed MyD88 alleles. Littermates were cohoused to ensure a shared microbiota. Each point represents an individual mouse. Error bars represent ±SEM. *P < 0.05; **P < 0.01; ns, not significant.