Fig. 2

mTORC2 negatively regulates NK cell cytotoxicity against tumor cells. a Flow cytometric analysis and cumulative frequencies of CD107a⁺ cells among total splenic NK cells (CD3⁻CD19⁻NK1.1⁺NKp46⁺) and within the indicated NK cell subpopulations from control versus Rictor^∆/∆ (top) or control versus Rictor^∆NK (bottom) mice after co-culture with Yac-1 targets in the presence of GolgiStop for 6 h. b The specific lysis of RMA-S tumor cells when co-cultured with NK cell subsets from control versus Rictor^∆/∆ or control versus Rictor^∆NK mice for 4 h at an E:T ratio of 1:1; NK cell subsets were sorted based on the expression of CD27 and CD11b, while RMA-S cells were labeled with Calcein-AM. c, d Flow cytometric analysis depicting perforin (c) and granzyme B (d) expression by splenic NK cells (CD3⁻CD19⁻NK1.1⁺NKp46⁺) and subpopulations thereof from control versus Rictor^∆/∆ (top) or control versus Rictor^∆NK mice (bottom) mice. e The cumulative ratio of RMA-S cell rejection 18 h after intraperitoneal injection of a 1:1 mixture of Cell Trace Far Red-labeled RMA and Cell Trace Violet-labeled RMA-S cells into control versus Rictor^∆/∆ or control versus Rictor^∆NK mice. f–h Flow cytometric analysis and cumulative results depicting expression of CD98 (f), CD71 (g), and the uptake of 2-NBDG (h) by total splenic NK cells or the indicated NK cell subsets, comparing control versus Rictor^∆/∆ (top) or control versus Rictor^∆NK (bottom) mice. The MFI was calculated relative to total NK cells or the CD27SP NK cell subset from control littermates. CD27SP, DP, and CD11bSP represent CD27⁺CD11b⁻, CD27⁺CD11b⁺, and CD27⁻CD11b⁺ NK cell subsets, respectively. For the bar graphs, each dot represents one mouse, and all experiments were replicated 5 (h bottom), 6 (a bottom, f, g, h top), 7 (c bottom, d bottom), or 8 (a top, c top, d top) times; error bars represent SD; ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ^∗∗∗∗p < 0.0001; unpaired two-tailed Student’s t test with Welsh’s correction (a, c, d, f–h); generalized linear models (b, e). Each of the shapes represents a matched pair of littermates in an individual experiment, and the horizontal red line represents the median (b, e)