Figure 2.

Id2 induces development of Lin⁻CD127⁺CD161⁺CD5⁺ cells from PNT CD34⁺CD1a⁻ cells. (A) Cell surface molecule expression on control or Id2-transduced cells after 7 days cultured on OP9 cells. (B) Fold expansion of control or Id2-transduced Lin⁻CD127⁺CD161⁺CD5⁺ and NK cells with or without IL-15. The cell number of control transduced Lin⁻CD127⁺CD161⁺CD5⁺ cells in the culture without IL-15 was set as 1. NK cells were determined by their cell surface expression of CD56 and low CD127. The data shown are an average of three independent experiments at day 13 of OP9 coculture. (C) Intracellular CD3 (clone OKT3) staining of Id2⁺Lin⁻CD127⁺CD161⁺ cells generated on OP9-Jag1 after 7 days. The population is further divided based on the expression of surface CD5 and α4β7. icCD3 histogram indicated in red: CD5⁺α4β7⁺, blue: CD5⁺α4β7⁻, black: CD5⁻α4β7⁻, gray filled: isotype control. (D) qPCR analysis of promyelocytic leukemia zinc finger (PLZF), Nfil3, and TOX mRNA expression in Id2⁺Lin⁻CD127⁺CD161⁺CD5⁺ cells generated on OP9–Jag1 and sorted at day 7 of coculturing. PNT CD34⁺CD1a⁻ cells and NK cells were used as negative and positive controls, respectively. All qPCR values presented are relative to GAPDH expression.