CCL3-EASER NK cell coculture controls. Cocultures presented in Fig. 7 were additionally incubated in the presence of monensin/brefeldin A to set Venus negative gates. (A–D) Representative contour plots of the coculture of EASER-NK cells with: (A) non-reporter NK/NKT cells; (B) OVA-activated CD4 OT-II T cells; (C) OVA-activated CD8 OT-I T cells; (D) thioglycolate elicited peritoneal macrophages. EASER-NK cells are indicated with an orange-shadowed gate, Ccl3-recipients with a gray-shadowed gate (exceptionally, recipient NK cells are indicated with a blue-shadowed gate, and NKT cells with a light-blue-shadowed gate), Venus⁺ cells with a green-shadowed gate, tdTomato⁺ cell with a red-shadowed gate, and tdTomato⁺Venus⁺ cells with a yellow-shadowed gate. (E) Representative contour plots of EASER-NK cells co-cultured with naive non-reporter NK cells. EASER-NK cells are indicated with an orange-shadowed gate, Ccl3-recipients with a gray-shadowed gate, Venus⁺ cells with a green-shadowed gate, tdTomato⁺ cell with a red-shadowed gate, and tdTomato⁺Venus⁺ cells with a yellow-shadowed gate. Data representative of three independent experiments (n = 4–5 per group). (F) Transwell assay of non-reporter (BL6) CD45.2 thioglycolate elicited macrophages (top chamber) and CD45.1 EASER-NK cells activated with plate bound anti-NK1.1 mAb (bottom chamber), panel 1. Macrophage Venus/tdTomato-negative control and Ccl3-Venus production control are shown in panels 2 and 3, respectively. Mac, macrophages; BFA, brefeldin A; Mon, monensin.