Effects of tumor-released extracellular nucleic acids on platelet and immune cell trafficking in experimental breast cancer. (A) Extracellular nucleic acids (red) released into the environment of 4T1 tumors as assessed by propidium iodide staining and in vivo microscopy on day 10 after tumor induction, a representative image (scale bar: 100 μm) and quantitative data for the fluorescence intensity relative to the distance from the tumor core (dotted line) are shown (mean ± SEM). (B) EC interactions of platelets in the peri- and intratumoral microvasculature of 4T1 tumors implanted into the left auricle in mice treated with blocking eTLR oligonucleotides or control oligonucleotides and of the left auricle of tumor-free mice; quantitative data are shown (mean ± SEM for n = 7 mice per group; #P < .05 vs control ∗P < .05 vs control oligo). Infiltration by (C) neutrophils, cMOs, B cells, CD8⁺ T cells, and (D) CD4⁺ T-cell subsets as assessed by flow cytometry as well as (E) size and weight of orthotopic 4T1 tumors, quantitative data are shown (mean ± SEM for n = 5 mice per group; ∗P < .05 vs control oligo). (F) Proliferation of 4T1 tumor cells exposed to agonists of TLR-7, -8, or -9 as assessed by 3-(4,5-dimethyldiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay and quantitative data are shown (mean ± SEM for n = 6 mice per group).