Figure 2. Gab3 is required for IL-2 and IL-15 induced priming and expansion of NK cells.

a) WT, Gab3^R27C and Gab3^KO NK cells stimulated with anti-NK1.1 antibody, YAC-1 cells or IL-12/IL-18 in the presence of IL-2. NK cell priming was measured by intracellular IFNγ staining (n=4 mice, ± s.e.m.). b-c) IL-2- or IL-15-induced NK cell expansion in vitro using WT, Gab3^R27C or Gab3^KO splenocytes (n=6, mean ± s.e.m.). Cells were gated on live NKp46⁺/NK1.1⁺ cells. d) Representative plots showing live WT, Gab3^R27C and Gab3^KO cell-trace violet-labeled splenocytes expanded with exogenous IL-2 or IL-15 in vitro for 5 days and stained for Ki67 (n=4, mean ± s.e.m.). e) Cell cycle analysis using EDU incorporation and 7AAD (DNA content) on IL-2- or IL-15-induced WT, Gab3^R27C and Gab3^KO NK cells (n=4, mean ± s.e.m.). Statistical analysis was done using a two-way ANOVA with Tukey post-test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.