Journal: Journal of Biological Chemistry

Article Title: Tumor Microenvironment Macrophage Inhibitory Factor Directs the Accumulation of Interleukin-17-producing Tumor-infiltrating Lymphocytes and Predicts Favorable Survival in Nasopharyngeal Carcinoma Patients

doi: 10.1074/jbc.m112.367532

Figure Lengend Snippet: FIGURE 3. Characterization and cytokine expression profiles of the Th17 cell subset among PBMCs and TILs from NPC patients. A, T cell surface markers were detected in Th17 cells from NPC patients. T cells were stimulated with phorbol 12-myristate 13-acetate/ionomycin for 4 h, then stained with fluorescence- conjugated antibodies against the markers shown, and analyzed by flow cytometry. B, graph of the percentage of cytokine-secreting Th17 cells among PBMCs and TILs from NPC patients (n 5) and PBMCs from healthy donors (n 5). Error bars represent the S.E. Significance was determined by the 2 test or paired t test. (*, p 0.001).

Article Snippet: Cytokines in the culture medium were measured using the Human Cytokine Array Panel A (Proteome Profiler; R&D Systems).

Techniques: Expressing, Staining, Fluorescence, Flow Cytometry

Journal: Journal of Biological Chemistry

Article Title: Tumor Microenvironment Macrophage Inhibitory Factor Directs the Accumulation of Interleukin-17-producing Tumor-infiltrating Lymphocytes and Predicts Favorable Survival in Nasopharyngeal Carcinoma Patients

doi: 10.1074/jbc.m112.367532

Figure Lengend Snippet: FIGURE 4. Generation and migration of Th17 cells mediated by NPC tumor cell lines. A, NPC tumor cell lines induce the differentiation of naive T cells into Th17cellsinvitro.PurifiedCD4naiveTcellsfromhealthydonorswerestimulatedwithOKT3andthenco-culturedwiththeirradiatedNPCcelllineC666(EBV) or CNE2 (EBV) in IL-2-containing medium for 7 days. The NP69 and LCL lines and the cytokines IL-1 and TGF were used as controls. All cultured cells were stained for Foxp3 and IL-17 for FACS analysis after stimulation of phorbol 12-myristate 13-acetate/ionomycin. The data represent one of three independent experiments. B, mean percentage of cytokine-secreting Th17 cells induced from CD4 naive T cells from three experiments. C, migration of Th17 cells was increased in response to cultured supernatants from the NPC cell lines C666 and CNE2, relative to that from the normal NP cell line NP69 or with medium alone. The data represent one of three independent experiments. D, mean percentage of cytokine-secreting Th17 cells in the inner well and outer well after migration induced by supernatants from NP69, C666, or CNE2 cells from three experiments. Error bars represent the S.E. Significance was determined by the 2 test or paired t test. (*, p 0.05).

Article Snippet: Cytokines in the culture medium were measured using the Human Cytokine Array Panel A (Proteome Profiler; R&D Systems).

Techniques: Migration, Cell Culture, Staining

Journal: British Journal of Cancer

Article Title: Impairment of carbonic anhydrase IX ectodomain cleavage reinforces tumorigenic and metastatic phenotype of cancer cells

doi: 10.1038/s41416-020-0804-z

Figure Lengend Snippet: a Proteome profiler Human XL Cytokine Array (ARY022, R&D Systems, MN, USA) probed with serum-free-conditioned medium (SF-CM) of C33a-FL-CA IX or C33a-NS-CA IX cells incubated under normoxic (NO) and hypoxic (HY) conditions for 48 h. Spot doublets of corresponding proteins with differential intensities are framed and indicated by arrows. Medium pooled from five independent experiments was used for the array. Spot intensities representing b pro-oncogenic basigin, and c anti-angiogenic thrombospondin-1 were analysed in culture medium samples of C33a-FL-CA IX versus C33a-NS-CA IX cells. Abundance of the proteins was assessed according to spot intensity using densitometry. Data are shown as mean ± SD. d Validation of the differential levels of basigin and thrombospondin-1, respectively, by Western blotting analysis of medium precipitates (MP) and cell lysates (CL) of normoxic (NO) and hypoxic (HY) cells. (* P < 0.05, ** P < 0.01, *** P < 0.005).

Article Snippet: PPA analyses were performed using the Human XL Cytokine Array Kit and Human Soluble Receptor Array Non-Hematopoietic panel kit (R&D Systems, MN, USA).

Techniques: Incubation, Biomarker Discovery, Western Blot

Journal: British Journal of Cancer

Article Title: Impairment of carbonic anhydrase IX ectodomain cleavage reinforces tumorigenic and metastatic phenotype of cancer cells

doi: 10.1038/s41416-020-0804-z

Figure Lengend Snippet: Analysis of the differential expression of a IGFBP-2 and b IGFBP-3 in SF-CM samples from C33a-FL-CA IX and C33a-NS-CA IX incubated under normoxic (NO) and hypoxic (HY) conditions for 48 h and evaluated by Proteome Profiler Human XL Cytokine Array (ARY022, R&D Systems, MN, USA). Abundance of the proteins was assessed according to spot intensity using densitometry. Data are shown as mean ± SD of five independent samples. c Western blotting analysis of CA IX, IGFBP-2 and IGFBP-3 protein levels in medium precipitates (MP) of C33a-FL-CA IX and C33a-NS-CA IX cells. Impairment of shedding in NS-CA IX is associated with increased IGFBP-2 and reduced IGFBP-3 secretion to culture medium. Quantitative PCR analysis of relative mRNA levels of d IGFBP-2, e IGFBP-3 and f IGF1 in C33a-FL-CA IX versus C33a-NS-CA IX cells normalised to β-actin mRNA. (*** P < 0.005, ns non-significant).

Article Snippet: PPA analyses were performed using the Human XL Cytokine Array Kit and Human Soluble Receptor Array Non-Hematopoietic panel kit (R&D Systems, MN, USA).

Techniques: Quantitative Proteomics, Incubation, Western Blot, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: IL-6R blockade with tocilizumab disrupts pericyte– and tumor cell–driven IL-6/STAT3 signaling, enhancing docetaxel efficacy in ER+ breast cancer

doi: 10.64898/2026.01.29.702661

Figure Lengend Snippet: a Cytokine expression. Fold change of cytokine array data highlighting IL-6, CXCL1, CXCL5, ST2, and GDF-15 from HBVP conditioned media. a.u. denotes arbitrary units. Data is representative of n = 2 independent experiments with technical duplicates. Mean ± s.e.m. shown. b IL-6 (pg/mL) from HBVP conditioned media quantified by IL-6 ELISA. Data is representative of n = 2 independent experiments with technical duplicates. Mean ± s.e.m. shown. c IL6 transcript levels in reference to GAPDH. a.u. denotes arbitrary units. Data is representative of n = 4 independent experiments with technical duplicates. Mean ± s.e.m. shown. P values were calculated by using an unpaired two-tailed t-test for a, b, and c . * P = 0.0245 ( a ), * P = 0.0134 ( b ), and * P = 0.0431 ( c ). * P ≤ 0.05. HBVP = human brain vascular pericytes. DTX = docetaxel.

Article Snippet: The conditioned media were collected after centrifuging for 5 minutes at RT and 600 g. The relative concentrations of 105 cytokines were measured using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, part of Bio-Techne #ARY022B).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Two Tailed Test