Journal: Oncology Letters
Article Title: Overexpression of B7-H4 promotes renal cell carcinoma progression by recruiting tumor-associated neutrophils via upregulation of CXCL8
doi: 10.3892/ol.2020.11701
Figure Lengend Snippet: Evaluation of the effects of CXCL8 on the B7-H4-mediated promotion of tumor growth and tumor-infiltrating neutrophils. 786-O transfectants were inoculated into non-obese diabetic/severe combined immunodeficiency mice (n=6/group). (A) Tumor growth curve was evaluated by assessing tumor volume. The tumor volumes on day 21 showed significance, ***P<0.001. (B) Tumor weight was evaluated after 21 days of treatment with isotype IgG or anti-CXCL8 antibody (n=6). (C) Fold-change of tumor weight between the isotype IgG and anti-CXCL8 antibody groups. (D) Gating strategy of CD11b + Ly6G + in tumor tissues is shown. (E) Tumor-infiltrating neutrophils were analysed by flow cytometry. Cells from tumor tissues were stained with fluorescein conjugated CD45, CD11b and Ly6G. The ratio of CD11b + Ly6G + in CD45 + cells was evaluated by flow cytometry. (F) Statistical analysis of the ratio CD11b + Ly6G + /CD45 + (n=6). Multiple comparisons among the groups were performed using the Tukey's post hoc test. *P<0.05, **P<0.01, ***P<0.001, as indicated. CXCL8, C-X-C motif chemokine ligand; B7-H4, B7 family member, H4; CD, cluster of differentiation; Ly6G, lymphocyte antigen 6 complex, locus G.
Article Snippet: The primary antibodies used were as follows: NME1 (1:500; cat. no. 11086-2-AP, Proteintech Group, Inc.); membrane metalloendopeptidase (MME; 1:500; cat. no 18008-1-AP; Proteintech Group, Inc.); vanin 1 (VNN1; 1:300; cat. no. 21745-1-AP; Proteintech Group, Inc.); matrix metalloproteinase 7 (MMP7; 1:300; cat. no. 10374-2-AP; Proteintech Group, Inc.); tumor necrosis factor α (TNF-α; 1:2,000; cat. no. 60291-1-Ig; Proteintech Group, Inc.); human C-X-C motif chemokine ligand (CXCL) 1/2/3 (1 μg/ml; cat. no. MAB276; R&D Systems, Inc.); CXCL8 (1:500; cat. no. 27095-1-AP; Proteintech Group, Inc.); human C-C motif chemokine ligand 20 (CCL20; 1 μg/ml); cat. no. MAB360; R&D Systems, Inc.); B7-H4 (1:500; cat. no. 12080-1-AP; Proteintech Group, Inc.); β-actin (1: 5,000; cat. no. 66009-1-Ig; Proteintech Group, Inc.) and α-tubulin (1:2,000; cat. no. 11224-1-AP; Proteintech Group, Inc.) were used.
Techniques: Flow Cytometry, Staining