PDK1 signaling is dispensable for NK cell terminal maturation and survival. (A) Representative flow cytometric profiles of NK1.1 versus NKp46 expression in splenic and BM CD3− cells from the indicated mice. Numbers show the percentages in square boxes among gated CD3− cells. (B and C) Flow cytometry analysis of NK1.1 versus CD122 expression in splenic CD3− cells in Rosa26 DTASTOP and Rosa26 DTASTOP/Ncr1-Cre+mice (left). Expression of CD27 versus CD11b on CD3−NK1.1+CD122+ NK cells was further analyzed (right). The numbers indicate the percentages of cells in each quadrant. (C) Absolute number of splenic CD3−CD122+NK1.1+ cells (left) and four-stage differentiating NK cell subsets (left), including DN, CD27SP, DP, and CD11bSP, were quantified in Rosa26 DTASTOP and Rosa26 DTASTOP/Ncr1-Cre+ mice. n = 4. ***, P < 0.0005. (D) Flow cytometry analysis of NK1.1 versus CD122 expression in CD3-negative splenic and BM cells of PDK1fl/fl and PDK1fl/fl/Ncr1-Cre+ mice. Numbers show the percentages in indicated circles among gated CD3− cells. (E) The absolute numbers of NK cells (CD3−NKp46+) in the spleens and BM of PDK1fl/fl and PDK1fl/fl/Ncr1-Cre+ mice were also quantified. Data were pooled from two independent experiments (n = 5–7). NS, not significant. (F) Representative flow cytometric profiles of the four-stage NK cell development, including CD27−CD11b−(DN), CD27+CD11b−(CD27SP), CD27+CD11b+(DP), and CD27−CD11b+(CD11b SP), from gated CD3−NK1.1+ NK cells in the spleens and BM of PDK1fl/fl and PDK1fl/fl/Ncr1-Cre+ mice are shown. The numbers indicate the percentages of cells in each quadrant.