Fig. 3

WEE1 kinase inhibition does not significantly alter KIL proliferation, viability or production of cytotoxic compounds. a KIL cells were cultured in AZD1775 (250 nM) or DMSO volume equivalent for 48 h in the presence or absence of IL-2 (20 ng/mL) and cell counts were obtained. b KIL cells were cultured in increasing doses of AZD1775 (250–2000 nM) or DMSO volume equivalent for 18 h and apoptosis was assessed by annexin V/7AAD flow cytometry. Representative dot plots shown on left, quantified for early (annexin V⁺7AAD⁻) or late (annexin V⁺7AAD⁺) apoptosis on right. * indicates significant difference compared to control. c KIL cells were cultured in AZD1775 (250 nM) or DMSO for 18 h and analyzed for expression of NKG2D by flow cytometry. Representative overlaid histograms on left, quantified on right. d KIL or sorted WT NK cells (stimulated with 20 ng/mL IL-2) were cultured in AZD1775 (250 nM) or DMSO for 18 h and analyzed for granzyme B accumulation via intracellular flow cytometry. Representative positive and isotype control dot plots of KIL cells shown on left, with KIL and sorted WT B6 NK staining quantified on the right. e KIL cells were combined with MOC2 cells at an E:T ratio of 3:1 for 4 h in the presence of brefeldin A, then KIL cells were assayed for IFNγ accumulation by intracellular flow cytometry. Representative positive and isotype control dot plots shown on left, quantified on the right. Results presented are representative of three independent experiments with similar results. *, p < 0.05. n/s, not significant