Innate CD8⁺ cells appearance in the thymus is a SP8 lineage decision.

CD45.1+ Control or T. cruzi-infected (Tulahuen) WT mice at day 12 post-infection (recipient mice) were anaesthetized and i.t. injected with 10 x 106 CD45.2+ thymocytes from the same control OT-I mouse (donor). After 48h, mice were sacrificed and the thymi were harvested. Dot plot shows the gate strategy for analysis of: (A) CD45.2+ SP8 OT-I thymocytes before being injected and after 48 h of being injected in (B) control or (C) T. cruzi-infected mice. Two days post i.t. injections (D) CD44, CD122, CD49d, Eomes and Tbet expression were analyzed by Flow cytometry in the CD45.2+ SP8 gated OT-I thymocytes. Changes in Eomes and Tbet levels were expressed as the difference between the MFI of Eomes or Tbet and the MFI from the corresponding Isotype control (IC). Histograms are representative of 2 independent experiments with 4–6 mice/group. The statistical test applied was a Student’s unpaired t test, Control vs T-cruzi *p.05 and **p.01.