Fig 3. iCD8α cells and osteopontin promote survival of NKp46⁺NK1.1⁺ IEL.

Enriched CD45⁺ IEL from small intestine and colon of Rag-2^-/- (a) or E8I^-/-Rag-2^-/- (b) mice were incubated in the presence or absence of recombinant osteopontin (2μg/ml final concentration). Cells were recovered 4 hours later, gated as described in the Materials and methods section, and analyzed for annexin V staining on gated NKp46⁺NK1.1⁺ IEL. Data are representative of at least two independent experiments (n = 4). (c) Enriched CD45⁺ cells from small intestine and colon of Spp-1^-/-Rag-2^-/- mice were incubated in the presence or absence of iCD8α cells derived from Rag-2^-/- mice, and analyzed as described above. In order to obtain enough iCD8α cells, 2–3 mice were pooled and counted as one sample (n = 4). Data is representative of at least 2 experiments. (d) Enriched CD45⁺ cells depleted from iCD8α cells from small intestine and colon of Spp-1^-/-Rag-2^-/- mice were incubated in the presence or absence of iCD8α cells derived from small intestine and colon of Rag-2^-/- or Spp-1^-/-Rag-2^-/- mice, and analyzed as described above. *p<0.05; ***p<0.001; ****p<0.0001 using unpaired two-tailed Student’s T test.