Journal: Cancer Growth and Metastasis

Article Title: Neutrophil-Derived Interleukin 16 in Premetastatic Lungs Promotes Breast Tumor Cell Seeding

doi: 10.1177/1179064417738513

Figure Lengend Snippet: IL-16 level is increased in lungs at a premetastatic stage. (A) Chemokine array performed on pooled total protein extracts obtained from premetastatic and control lungs (n = 5) and dot quantification (right panel). (B) IL-16 measurement by ELISA performed on lungs protein extracts. (C) Representative Western blot of secreted IL-16 production at day 7 in lungs of tumor-bearing or control mice (blot performed on pooled total protein extracts obtained, n = 9-10). Actin serves as a loading control. ELISA anti-IL-16 performed on (D) serum samples and (E) bronchoalveolar lavage fluids obtained from tumor-bearing mice or control mice at day 7. Results are expressed as mean ± SEM. ** P < .01, Student t test (n = 10). (F) Representative immunohistochemistry against IL-16 performed on lung sections obtained from tumor-bearing and control mice sacrificed at days 7 and 21 following the primary tumor implantation. Scale bar represents 200 µm. ELISA indicates enzyme-linked immunosorbent assay; IL-16, interleukin 16.

Article Snippet: IL-16 detection in lungs, serum samples, and BALF and detection of s100A8, KC, and GM-CSF in 4T1-conditioned medium were performed with respective Mouse DuoSet ELISA kits (R&D Systems) according to the manufacturer’s protocol.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Western Blot, Immunohistochemistry, Tumor Implantation

Journal: Cancer Growth and Metastasis

Article Title: Neutrophil-Derived Interleukin 16 in Premetastatic Lungs Promotes Breast Tumor Cell Seeding

doi: 10.1177/1179064417738513

Figure Lengend Snippet: Neutrophils expressed IL-16 in premetastatic lungs. (A) Representative immunofluorescence experiments showing a colocalization between neutrophil foci and IL-16–positive area in premetastatic lungs. Scale bar corresponds to 100 µm. (B) Representative flow cytometry plots showing the purity of neutrophil isolation by MACS technologies. (C) Representative cytocentrifugation of neutrophils obtained using MACS technologies. Scale bar represents 100 µm. (D) IL-16 dosage by enzyme-linked immunosorbent assay in culture supernatant of neutrophils treated or not with 4T1-conditioned medium. Results are expressed as mean ± SEM. ** P < .01, Student t test. (E) Neutrophil percentage in bronchoalveolar lavage of tumor-bearing mice treated with a Ly6G blocking antibody or a control isotype. Results are expressed as mean ± SEM. ** P < .01, Mann-Whitney test (n = 6). (F) IL-16 dosage in lungs of tumor-bearing mice treated with a Ly6G blocking antibody or a control isotype. Results are expressed as mean ± SEM. * P < .05, Student t test (n = 6). IL-16 indicates interleukin 16; MACS, magnetic-activated cell sorting.

Article Snippet: IL-16 detection in lungs, serum samples, and BALF and detection of s100A8, KC, and GM-CSF in 4T1-conditioned medium were performed with respective Mouse DuoSet ELISA kits (R&D Systems) according to the manufacturer’s protocol.

Techniques: Immunofluorescence, Flow Cytometry, Isolation, Enzyme-linked Immunosorbent Assay, Blocking Assay, Control, MANN-WHITNEY, FACS

Journal: Cancer Growth and Metastasis

Article Title: Neutrophil-Derived Interleukin 16 in Premetastatic Lungs Promotes Breast Tumor Cell Seeding

doi: 10.1177/1179064417738513

Figure Lengend Snippet: Soluble factors derived from 4T1 cells induced IL-16 production in the pulmonary microenvironment. (A) ELISA against IL-16 on whole lung protein homogenates obtained from mice intratracheally instilled with control medium or 4T1-conditioned medium. Results are expressed as mean ± SEM. * P < .05, Mann-Whitney test (n = 6). (B) Percentage of cell content in BAL of mice intratracheally instilled with control medium or 4T1-conditioned medium (EC: epithelial cells, NEU: neutrophils, EO: eosinophils, LT: lymphocytes, and MP: macrophages). Results are expressed in mean ± SEM. * P < .05, Student t test (n = 6). (C) Representative Xenogen IVIS analysis and bioluminescence quantification of lungs obtained from mice treated with control medium or 4T1-conditioned medium and receiving an intravenous injection of 4T1 cell. Results are expressed as mean ± SEM. * P < .05, Student t test (n = 6). IL-16 indicates interleukin 16.

Article Snippet: IL-16 detection in lungs, serum samples, and BALF and detection of s100A8, KC, and GM-CSF in 4T1-conditioned medium were performed with respective Mouse DuoSet ELISA kits (R&D Systems) according to the manufacturer’s protocol.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Control, MANN-WHITNEY, Injection