Figure 3.

Eomes^lo and Eomes^hi subsets correspond to different cell lineages. (A) Scheme of adoptive transfer of Eomes^hi cells into CD45.1 mice bearing cancer (relative to mock). After injection of B16F10 cells (day 0), Eomes^hi cells were harvested from donor Eomes-GFP mice and adoptively transferred into recipient mice at day 4. The mice were then sacrificed at day 10 and lungs were harvested and analyzed. (B) Eomes^lo and Eomes^hi cells in Eomes-GFP mice (Red) overlaid with Eomes FMO (blue) (left panel). Two distinct populations can be seen upon running live cells under flow cytometer; Sorting efficiency ~99% middle panel); Donor cells after transfer of CD45.2⁺ Eomes^hi-GFP⁺ cells into CD45.1 mice (right panel). (C) Eomes (MFI) after transfer of donor derived Eomes^hi GFP⁺ cells in cancer-bearing (B16F10) and cancer-lacking (mock) hosts at day 10. No significant difference in Eomes MFI was observed between cancer and mock mice. (D) Frequency of Eomes^lo GFP⁻ cells amongst donor NK cells at day 10 post-injection of B16F10 cells (E) Flow plots and graphs show near absence of Eomes^lo cells in T-bet KO mice (~0.8%) compared to WT mice (~29.1%), indicating that T-bet is needed for development of Eomes^lo cell development (Red—NKp46⁺NK1.1⁺ cells; Blue—Eomes FMO). MFI is Mean Fluorescence Intensity, n = 3–4 for each group. Results are representative of three independent repeats; data are presented as mean ± s.e.m.; Significance was tested using two-tailed students' t-test.