Fig. 3

DHA feeding improved IFN-γ production and degranulation by NK cells in the spleen at day 7 postinfection. A Flow cytometric analysis of the ratio and the geometric mean fluorescence intensity (gMFI) of IFN-γ⁺ splenic NK cells (CD3⁻CD19⁻NK1.1⁺NKp46⁺) from control versus DHA-fed mice following stimulation with PMA and ionomycin in the presence of GolgiPlug and GolgiStop for 6 h. B, C Flow cytometric analysis depicting the frequencies and gMFI of perforin (B) and granzyme B (GZMB) (C) by splenic NK cells (CD3⁻CD19⁻NK1.1⁺NKp46⁺) from control versus DHA-fed mice at day 7 postinfection. D Purified splenic NK cells from both DHA-fed and control mice at day 7 after MCMV infection were cocultured with CTV-labeled YAC-1 (left) or MCMV-infected 3T3 cells (right) for 6 h at the E:T ratio of 16:1. Representative flow cytometry plots of CTV⁺ singlets are shown on the left. The percentages of NK cell-specific lysis are shown on the right, which were calculated by the following formula: [(% CTV ⁺ 7-AAD⁺ cell specific lysis − %CTV ⁺ 7-AAD⁺ cell spontaneous lysis)/ (100 − % CTV ⁺ 7-AAD⁺cell spontaneous lysis)] × 100. For each experiment, n = 9 pooled from 3 independent experiments (A–C); n = 5 pooled from 2 independent experiments (D). Each symbol represents an individual mouse, and the blue dots and red square represent control and DHA-enriched diet-fed mice, respectively. All experiments were replicated 3 times. Error bars represent interquartile ranges; *p < 0.05; **p < 0.01