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: Virus research

Article Title: Paracrinal regulation of neutrophil functions by coronaviral infection in iPSC-derived alveolar type II epithelial cells.

doi: 10.1016/j.virusres.2024.199391

Figure Lengend Snippet: Fig. 5. Identification of cytokines secreted by infected iAECIIs and their effect on HL-60 cells. (A) Cytokine array analysis showing the levels of the indicated cy tokines in the medium conditioned by iAECIIs infected with HCoV-229E for 24 or 48 h (24 hpi and 48 hpi). Ctrl – uninfected control. Right panel: cytokine label legend. (B) Western blot analysis of the expression of IL-8 and ICAM-1 in HCoV-229E-infected iAECIIs. (C) Transwell migration assay showing the migration capacity of HL-60 neutrophils in response to a medium conditioned by iAECIIs infected with HCoV-229E for 48 h (48 hpi) in the absence or presence of an IL-8-neutralizing antibody (IL-8 nAb). Ctrl – uninfected control conditioned medium. Data shown as means with SD error bars, n = 3, *p < 0.005, (ANOVA). (D) Representative fluorescence microscopy images of adherent HL-60 neutrophils after incubation with infected iAECIIs in the absence or presence of an ICAM-1-neutralizing antibody (ICAM-1 nAb). Ctrl – uninfected control. (E) Cell count of adherent HL-60 neutrophils. Mean values are shown with SD error bars, n = 3, *p < 0.005.

Article Snippet: Cytokines in the conditioned medium were detected using the Human Cytokine Array Kit (#ARY005B; R&D Systems), and the test strips were moistened and activated according to the manufacturer’s user manual.

Techniques: Infection, Control, Western Blot, Expressing, Transwell Migration Assay, Migration, Fluorescence, Microscopy, Incubation, Cell Counting

Journal: Virus research

Article Title: Paracrinal regulation of neutrophil functions by coronaviral infection in iPSC-derived alveolar type II epithelial cells.

doi: 10.1016/j.virusres.2024.199391

Figure Lengend Snippet: Fig. 6. RNA-seq analysis identifies upstream cytokine pathways in the immune responses by neutrophils. (A and B) Cytoscape ClueGO networks of upregulated genes by infection of iAECIIs with HCoV-229E (A) or triggered by infected HCoV-229 conditioned medium in HL-60 cells (B). (C and D) Bubble plots showing the most enriched GO-BP terms among the genes upregulated by the infection of iAECIIs with HCoV-229E (C) or the genes upregulated in HL-60 cells by cultivation in the infected HCoV-229-conditioned medium (D). (E and F) Hierarchical clustering heatmaps showing the signatures of genes differentially regulated in infected iAECIIs (E) and conditioned medium-stimulated HL-60 cells (F). (G and H) X2K network analysis showing the kinases and transcription factors predicted to regulate differentially expressed genes in infected iAECIIs (G) and conditioned medium-stimulated HL-60 cells (H).

Article Snippet: Cytokines in the conditioned medium were detected using the Human Cytokine Array Kit (#ARY005B; R&D Systems), and the test strips were moistened and activated according to the manufacturer’s user manual.

Techniques: RNA Sequencing, Infection

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

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: High CX3CL1 enrichment in the CI group predicts poor prognosis of GC patients. ( A ) Schematic of detecting cytokines in mouse serum via cytokine array. ( B ) Volcano plots depicting the differential analysis of detected cytokines in CI and CON( n = 3 per group). ( C ) Schematic of screening differentially expressed cytokines between CI and CON groups by P value and log 2 (FC). ( D , E , F ) Survival curves for GC patients with low or high expression of MMP2/COL18A1/CX3CL1 in GEO database using Kaplan-Meier method. ( G ) The mRNA level of CX3CL1 in GC tissues compared with normal tissues in TCGA-STAD datasets. ( H ) The mRNA level of CX3CL1 in GC tissues between different T-stage from TCGA-STAD datasets. ( I ) Survival curves for GC patients with low or high expression of CX3CL1 in TCGA database using Kaplan-Meier method. ( J ) Concentration of CX3CL1 in the mice tumors detected by ELISA( n = 3 per group). Data are represented as mean ± SEM. * P < 0.05,** P < 0.01,*** P < 0.001(J: two-tailed unpaired Student’s t test).

Article Snippet: The concentrations of CX3CL1 in the tumor tissue were quantified using commercially CX3CL1 ELISA kit (Proteintech, KE10076) according to the kit instructions.

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

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: Single-cell transcriptomic characterization of CX3CL1 expression. ( A ) Dimensionality reduction and sample visualization analysis of tSNE. ( B ) Manual annotation of cell types based on Marker gene annotation. ( C ) tSNE plots of scRNA-seq from CON and CI groups. ( D ) Inflammation score mapping illustrating differences in the cellular distribution of inflammatory transcriptional signatures between CON and CI tumors. ( E ) Violin plot showing the distribution of Cx3cl1 expression across annotated cell types. ( F ) Feature plot visualization of Cx3cl1 expression on the t-SNE map in CON and CI groups. ( G ) Violin plot showing the distribution of Cx3cr1 expression across annotated cell types. ( H ) Feature plot visualization of Cx3cr1 expression on the t-SNE map in CON and CI groups. ( I ) GO enrichment analysis. ( J ) KEGG enrichment analysis of differentially expressed genes comparing the Tumor_6 subcluster with other tumor cell subclusters. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The concentrations of CX3CL1 in the tumor tissue were quantified using commercially CX3CL1 ELISA kit (Proteintech, KE10076) according to the kit instructions.

Techniques: Single Cell, Expressing, Marker

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: CX3CL1 promotes gastric cancer cell proliferation and migration via CX3CR1. ( A , B ) Relative mRNA expression levels of CX3CR1/Cx3cr1 in GES-1, HGC-27, RAW264.7 and MFC cells. ( C ) Protein expression of CX3CR1 in GES-1, HGC-27, RAW264.7, and MFC cells. ( D , E ) Cell viability of MFC ( D ) and HGC-27 ( E ) cells treated with increasing concentrations of CX3CL1 ( n = 3 per group). ( F , G ) Representative images ( F ) and quantification ( G ) of colony formation assays in MFC and HGC-27 cells treated with CX3CL1( n = 3 per group). ( H , I ) Representative images ( H ) and quantification ( I ) of Transwell migration assays in MFC and HGC-27 cells treated with CX3CL1 ( n = 3 per group). Scale bar, 50 μm. ( J , K ) Representative images ( J ) and quantification ( K ) of wound-healing assays in MFC and HGC-27 cells treated with CX3CL1 for 24 h ( n = 3 per group). Scale bar, 500 μm. ( L - P ) The effects of CX3CL1 on cell viability ( L ), wound-healing migration ( N ), and Transwell migration ( O , P ) were further examined in the presence or absence of the CX3CR1 inhibitor JMS-17-2. Data are presented as mean ± SEM from independent experiments. For panels D and E, statistical significance is indicated relative to the 0 ng/mL CX3CL1 group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant. ( D , E ) one-way ANOVA with Dunnett’s multiple comparisons test; ( F - K ) two-tailed unpaired Student’s t-test; (L - P) two-way ANOVA with Sidak’s multiple comparisons test(interaction P = 0.0034, 0.0079, 0.0033, 0.0163, 0.0156, and 0.0311 for panels L - P , respectively).

Article Snippet: The concentrations of CX3CL1 in the tumor tissue were quantified using commercially CX3CL1 ELISA kit (Proteintech, KE10076) according to the kit instructions.

Techniques: Migration, Expressing, Two Tailed Test

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: ADAM10-mediated increased cleavage of CX3CL1 contributes to CI-promoted GC progression. ( A-C ) RT-qPCR and Western blot confirmed that CI increased the mRNA and protein expression of ADAM10 in tumor tissues( n = 3 per group). ( D ) Immunofluorescence assay suggested the increase of ADAM10 (Red) fluorescence intensity in tumor tissues. ( E-F ) Western blot confirmed that LPS increased the protein expression of ADAM10 in GC cell lines( n = 3 per group). ( G ) ELISA assay demonstrated that the ADAM10 antagonist GI254023X could attenuate the LPS-induced increase in soluble CX3CL1 levels in GC cells( n = 3 per group). ( H-N ) ADAM10 antagonist GI254023X inhibited LPS-promoted GC cells migration and invasion( n = 3 per group). ( O ) Schematic diagram shows the experimental protocol for determine the effect of ADAM10 inhibitor GI in suppressingsing CI-promoted tumor progression in mice( n = 5 per group). ( P-Q ) Left: Representative macroscopic images of tumors; Right: weight of tumors( n = 5 per group). ( R ) Concentration of CX3CL1 in the mice tumors detected by ELISA( n = 3 per group). Data are represented as mean ± SEM. For panel F, statistical significance was determined relative to the 0 µg/mL LPS group. For panels G , I , K , N , statistical significance was determined relative to the LPS group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001(A, C: two-tailed unpaired Student’s t test; F , G , I , K , N , Q , R : one-way ANOVA with Dunnett’s multiple comparisons test).

Article Snippet: The concentrations of CX3CL1 in the tumor tissue were quantified using commercially CX3CL1 ELISA kit (Proteintech, KE10076) according to the kit instructions.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay, Migration, Concentration Assay, Two Tailed Test

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: Graphical Abstract. CI induces the upregulation of ADAM10 expression in GC cells. Upregulated ADAM10 mediates the cleavage of membrane-bound CX3CL1 (mCX3CL1), thereby promoting the release of soluble CX3CL1 (sCX3CL1). The increased sCX3CL1 ultimately enhance GC cell proliferation and migration to accelerate tumor progression.

Article Snippet: The concentrations of CX3CL1 in the tumor tissue were quantified using commercially CX3CL1 ELISA kit (Proteintech, KE10076) according to the kit instructions.

Techniques: Expressing, Membrane, Migration