Splenic NKp46highNK cells produced the highest levels of TNF-α. Analysis of TNF-α production of NKp46-defined NK-cell subsets (CD8α+NKp46-: blue, CD8α+NKp46+: green, CD8αdim/-NKp46high: red) isolated from blood and spleen. FACS-sorted CD3-CD8α+NKp46- and CD3-CD8α+NKp46+ NK cells from blood and CD3-CD8α+NKp46-, CD3-CD8α+NKp46+ and CD3-CD8αdim/-NKp46high NK cells from spleen were stimulated with rhIL-2, rpIL-12 and rpIL-18 (A) or PMA and Ionomycin (B) for 24 h. Supernatants were tested for TNF-α production in ELISA. Bar graphs on the left show data from one representative animal and are displayed as the mean of duplicates ± SD. TNF-α production of four animals analysed are shown on the right. Mean values are represented by a black bar. Significant differences between the subsets in blood or spleen are indicated (* = p < 0.05, ** = p < 0.01).