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. 2019 Sep 4;10:2088. doi: 10.3389/fimmu.2019.02088

Figure 2.

Figure 2

4T1 tumor-bearing mice but not 67NR tumor-bearing mice exhibit neutrophilia and a NET-dependent prothrombotic state. (A) Systemic levels of G-CSF in control (gray bars, n = 5), 67NR (open bars, n = 3) and 4T1 (black bars, n = 5) tumor-bearing mice were assessed by a specific ELISA kit as described in section Materials and Methods. (B) Neutrophil counts in control (gray bars, n = 12), 67NR (open bars, n = 5) and 4T1 (black bars, n = 11) tumor-bearing mice were evaluated as described in section Materials and Methods. Two systemic markers of NETs, MPO (C) and cfDNA (D), were analyzed in control (gray bars, n = 5), 67NR (open bars, n = 3) and 4T1 (black bars, n = 6) tumor-bearing mice as described in section Materials and Methods. For systemic analysis, the blood was collected 21 days after the tumor cell inoculation. (E) Venous thrombosis was evaluated with stasis of the inferior vena cava for 6 h in control (•, n = 14), 67NR (▴, n = 14), and 4T1 (■, n = 15) tumor-bearing mice. (F) Alternatively, 4T1 tumor-bearing mice (■, n = 12) were treated with DNase I (50 μg, i.v.) (▴, n = 7) or GSK484 (10 mg/kg, i.v.) (▾, n = 9) just before performing the procedure to induce stasis of the inferior vena cava for 3 h. Each dot represents one individual mouse. Values represent the mean ± SD. *P < 0.05, **P = 0.01, and ***P = 0.001; n.s., non-significant, analysis of variance (ANOVA) followed by Tukey's post-hoc test.