Fig 1: CCL17 expression in clear cell renal cell carcinoma (ccRCC) tissues and Smooth estimates of HR (+1 IOD). Representative CCL17 immunohistochemical (IHC) images of ccRCC tumor tissues with low CCL17 expression (a) and high CCL17 expression (b). Smooth estimates of HR (+1 IOD) showed a higher risk of death or recurrence for patients with lower CCL17 expression (c), (e). Smooth estimates of HR (using IOD = 8461 as a reference) showed a significant and stable prognostic difference between patients with high/low CCL17 expression (d), (f). Dashed lines: 95% confidence bands
Fig 2: CCL17 derived from lactate-activated TAMs mediates the invasion of PAs through binding to CCR4. A-C, Quantification of various TAM-derived chemokines and cytokines in PMA-treated THP1 cells stimulated with lactic acid (A), CM from human primary PA cells (B), or IL4 (10 ng/ml) (C) for 24 h. D, Representative images of wound-healing assay using GH3 cells in the presence or absence of CCL17 (50 ng/ml), AZD2098 (20 µM), or combination. E, PCNA and EMT biomarker protein expression in GH3 cells under stimulation with CCL17, AZD2098, or combination for 24 h. F, Proliferation of GH3 cells following 24, 48, and 72 h stimulation under CCL17, AZD2098, or combination. G, Cell cycle assays in GH3 cells in the presence or absence of CCL17, AZD2098, or combination. H, Tumor growth curve in tumors derived from NOD/SCID tumor-bearing mice treated with PBS, AZD2098 (1.5 mg/kg), CCL17 (0.1 µg/kg), or combination. I, Representative images and J, mass weight of tumors derived from NOD/SCID tumor-bearing mice treated with PBS, AZD2098, CCL17, or combination. K, HE staining of edge region of tumor and muscle tissues derived from the PBS, AZD2098, CCL17, or combination group. All t-tests were two-tailed. Mean ± SEM. *P < 0.05, **P < 0.01, and ***P < 0.001.
Fig 3: CircCHST15 regulated the tumor growth, the T cell subtype, and the secretion of related-factors in mouse tumor tissues. (A) The solid tumors were shown. (B) The weight of a solid tumor was calculated. (C, D) The level of CD4+ T cells in the blood (C) and tumor supernatant (D) of mice was quantified by the flow cytometer. (E, F) The level of CD8+ T cells in the blood (E) and tumor supernatant (F) of mice was quantified by the flow cytometer. (G, H) The level of Tregs in the blood (G) and tumor supernatant (H) of mice was calculated based on the formula: Tregs+ (%) = [(CD3+ + CD4+ + CD25+ + total cell number)/CD3+ + total cells number] × 100%. (I, J) The proportion of immune cell subsets in peripheral blood and tumor was measured by flow cytometry. (K–M) The levels of IFN-? (K), TNF-ß (L), and IL-10 (M) in the blood of mice were detected by ELISA. (N, O) The expressions of CCL17 and CCL22 in the mouse tumor tissues were detected by Western blot, GAPDH was used as an internal control. (+++ P < 0.001, vs. NC; **P < 0.01, ***P < 0.001, vs. Control; ^ P < 0.05, ^^ P < 0.01, ^^^ P < 0.001, vs. mock; ### P < 0.001, vs. CircCHST15; & P < 0.05, &&& P < 0.001, vs. Anti-PD-L1).
Fig 4: Basic physiological indicators and cardiac functional biomarkers after the injection of CCL17 neutralizing antibody in mice. 8–12-wk-old C57BL/6J background WT mice were treated with IgG control or anti-CCL17 antibody 28 d after saline or Ang II infusion. (A and B) Systolic and diastolic blood pressures (A) and heart rates (B) in C57BL/6J background WT mice treated with IgG control or anti-CCL17 antibody after saline or Ang II infusion (n = 12–13 biological replicates). (C) qRT-PCR was performed to analyze the mRNA levels of cardiac function (Anp, Bnp), hypertrophic (ß-Mhc, Acta1, Rcan1.4), and fibrosis (Ctgf, Col1a1, Col3a1) marker (n = 6 biological replicates). Data shown in A–C are presented as the mean ± SD. Statistical significance was tested using one-way ANOVA followed by Bonferroni post hoc test (A and B). Unpaired Student’s t-test was used for equal variance or Welch t-test used for unequal variance (C). *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Fig 5: Accumulation of CD45+ cells, CD4+ T cells, macrophages, DCs, and neutrophils within the cardiac tissue of WT mice after infusion with Ang II for 4 wk. (A) Immunofluorescence analysis of CD4+ T cells (anti-CD4), macrophages (anti-F4/80), DCs (anti-CD11c), and neutrophils (anti-Ly6G) expression within the cardiac tissue of 8–12-wk-old C57BL/6J background WT mice after infused with Ang II for 4 wk (n = 6 biological replicates). Scale bars: 20 µm. (B and C) Flow cytometry analysis of CD45+ T cells (B) and CD3+CD4+ T cells (C) of 8–12-wk-old C57BL/6J background WT and Ccl17-KO mice treated with saline or Ang II for 4 wk in heart tissues (n = 6 biological replicates). All data are presented as the mean ± SD. Statistical significance was tested using one-way ANOVA followed by Bonferroni post hoc test (A–C). **, P < 0.01; ***, P < 0.001.
Supplier Page from Abcam for Anti-TARC/CCL17 antibody