Fig. 5

Ackr2^−/− neutrophils have increased tumor-killing activity and expression of inflammatory CC chemokine receptors. a MFI of CellROX emission by WT and Ackr2^−/− neutrophils. Data are normalized on MFI of WT neutrophils (n = 9 for WT and 5 for Ackr2^−/−). b qPCR analysis of chemokine receptors and c activation markers in FACS-sorted neutrophils taken from unchallenged WT (white columns) and Ackr2^−/− (black columns) mice (n = 4 for both WT and Ackr2^−/− mice, two independent experiments). Data are relative to Gapdh expression. d Percentage of WT and Ackr2^−/− CD45.2 neutrophils on total CD45.2 transferred neutrophils in the lungs of WT CD45.1 hosts and e absolute number of circulating WT and Ackr2^−/− CD45.2 neutrophils after 1 h from adoptive transfer in CD45.1 hosts and i.p. injection of CCL3L1 (n = 3 recipients for each group). f CellROX MFI in WT (white columns) and Ackr2^−/− (black columns) neutrophils preincubated with PBS or LPS (100 ng/ml, 20 min) and stimulated with CCL2 (500 ng/ml, 30 min) or PMA (50 ng/ml, 30 min). CellROX MFI was normalized on basal WT group (n = 4, two independent experiments for both WT and Ackr2^−/− mice). g 4T1-luc cell killing by magnetically sorted circulating neutrophils taken from tumor-bearing WT (white columns) and Ackr2^−/− (black columns) mice 21 days after 4T1 injection. Cells were treated with medium or LPS (100 ng/ml) + PMA (50 ng/ml). Where indicated, apocynin (100 μM) was added (n = 3, two independent experiments for both WT and Ackr2^−/− mice). Data are represented as mean (SD). p value was generated using the unpaired t-test. *p < 0.05, **p < 0.01, ***p < 0.001