Journal: Molecular medicine reports

Article Title: Analysis of murine and human Treg subsets in inflammatory bowel disease.

doi: 10.3892/mmr.2017.6912

Figure Lengend Snippet: Figure 5. Serum inflammatory cytokines in patients with ulcerative colitis. Serum from ulcerative colitis patients and healthy donors were collected and inflammatory cytokines were determined by ELISA. *P<0.05, **P<0.01, ***P<0.001 and ****P<0.0001 as indicated. IL, interleukin; TSLP, thymic stromal lymphopoietin; MMP, matrix metalloproteinase.

Article Snippet: Serum cytokine levels were analyzed using the following commercially available ELISA kits according to the manufacturer's protocol: MMP-2, Total MMP-2 Quantikine ELISA kit (cat. no. MMP200; Quantikine, R&D Systems, Inc., Minneapolis, MD, USA); thymic stromal lymphopoietin, Legend MaxTM Human TSLP ELISA kit with Pre-coated Plates (cat. no. 434208; BioLegend, Inc., San Diego, CA, USA); MMP-9, Legend MaxTM Human MMP-9 ELISA kit with Pre-coated Plates (cat. no. 440707; BioLegend, Inc.); IL-17A, Legend MaxTM Human IL-17A ELISA kit with Pre-coated Plates (cat. no. 433918; BioLegend, Inc.); IL-25, Human IL-17E/IL-25 AccuSignal ELISA kit (cat. no. KOA0468; Rockland Immunochemicals, Inc., Limerick, PA, USA); and eotaxin, Human Eotaxin ELISA kit (cat. no. KOA0159; Rockland Immunochemicals, Inc.).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Macrophage-derived CCL20 promotes abdominal aortic aneurysm progression via lymphocytes CCR6

doi: 10.3389/fimmu.2026.1780720

Figure Lengend Snippet: Macrophages Exhibit Altered Gene Expression and Secrete Abundant CCL20. (A) The volcano plot of DEGs of macrophages in the normal and AAA groups in GSE166676 and GSE226492 . (B) Bubble plot of GO BP enrichment results for DEGs. (C) Bubble plot of enriched KEGG pathways for DEGs. (D) GSEA results based on KEGG identified activated and suppressed pathways of macrophages in the normal and AAA groups. Categories with red dots (NES > 0) showed the activated pathways in AAA group, while categories with blue dots (NES < 0) showed the activated pathways in the normal group. (E) The mRNA expression of IL-1beta, IL-8, TNF-α in normal and AAA tissues from human. (F) The mRNA expression of ACC1, FASN, and PPAR-γ in human Normal and AAA groups. (G) Representative images of Oil red O staining of the normal (n=5) and AAA group (n=6). Scale bar:10, 25μm. (H) All up DEGs in the Cytokine-Cytokine receptor interaction pathway. (I) CCL20 expression in 11 cell types. (J) Representative images of IF co-localization of CCL20 (green) and macrophages (CD68, red). Nuclei were stained with DAPI (blue), scale bar, 50 μm. The synchronized fluctuations of CCL20 (green) and macrophages (CD68, red) curves indicated co-localization at these discrete sites. (K) CCL20 expression in 11 cell types between the normal group and AAA groups. (L) ELISA test of CCL20 in the normal group (n=79) and AAA group (n=80). (M) ROC curve comparing models for AAA diagnosis. The model achieved an area under the curve (AUC) of 0.7. (N) Representative images of IF of CCL20 (green) expression in the the normal (n=5) and AAA group (n=6). Nuclei were stained with DAPI (blue), scale bar, 50 μm. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Serum CCL20 levels in AAA patients and healthy controls were quantified using a commercial ELISA kit (Proteintech, KE00149, China), following the manufacturer’s protocol.

Techniques: Gene Expression, Expressing, Staining, Enzyme-linked Immunosorbent Assay, Biomarker Discovery

Journal: Journal of Neuroinflammation

Article Title: Astroglial TLR9 antagonism promotes chemotaxis and alternative activation of macrophages via modulation of astrocyte-derived signals: implications for spinal cord injury

doi: 10.1186/s12974-020-01748-x

Figure Lengend Snippet: ODN 2088 modulates the release of chemokines by SC astrocytes, in vitro. a Representative chemokine arrays used to detect chemokines in CM of vehicle- and ODN 2088-treated astrocytes. The chemokine arrays were independently repeated twice, showing similar results. Results from a representative experiment are shown. The dots enclosed in rectangular boxes show chemokines whose levels were decreased (> 5% difference) in the CM of ODN 2088-treated astrocytes compared to the CM of vehicle-treated astrocytes. The dots enclosed in the oval box show the chemokine whose levels were increased (> 5% difference) in the CM of ODN 2088-treated astrocytes compared to CM of vehicle-treated astrocytes. 1: CCL1; 2: CCL9/MIP-1γ; 3: CCL2/MCP-1; 4: CCL20/MIP-3α; 5: CX3CL1. b Densitometric quantification of the signal obtained in the chemokine array using the Image Lab software (Bio-Rad). c Quantification of CCL9 levels in CM obtained from ODN 2088- or vehicle-treated astrocytes [** p < 0.01, independent-sample t -test, two-tailed]. The experiment was independently repeated four times, and the mean of 4 experiments ( n = 4) is shown. d Quantification of CCL2 levels in CM obtained from ODN 2088- or vehicle-treated astrocytes [* p < 0.05, independent-sample t -test, two-tailed]. The experiment was independently repeated three times, and the mean of 3 experiments ( n = 3) is shown

Article Snippet: In addition, CCL1 concentrations were independently measured by a second ELISA kit purchased from a different vendor (Rockland Immunochemicals; Limerick, PA, USA).

Techniques: In Vitro, Software, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: Astroglial TLR9 antagonism promotes chemotaxis and alternative activation of macrophages via modulation of astrocyte-derived signals: implications for spinal cord injury

doi: 10.1186/s12974-020-01748-x

Figure Lengend Snippet: CCL1 released by ODN 2088-treated astrocytes mediates the chemotaxis of peritoneal macrophages. Quantification of F4/80 + cells that crossed to the lower surface of the membrane in response to CM derived from vehicle- or ODN 2088-treated TLR9 astrocytes in the absence or presence of CCL1 neutralizing antibody or IgG 2A isotype control [ F (5, 48) = 81.03, p < 0.0001 by one-way ANOVA, **** p < 0.0001 by Tukey’s post hoc test]. The results of three independent experiments ( n = 3) are shown. Data are presented as mean ± SEM

Article Snippet: In addition, CCL1 concentrations were independently measured by a second ELISA kit purchased from a different vendor (Rockland Immunochemicals; Limerick, PA, USA).

Techniques: Chemotaxis Assay, Membrane, Derivative Assay, Control

Journal: Journal of Neuroinflammation

Article Title: Astroglial TLR9 antagonism promotes chemotaxis and alternative activation of macrophages via modulation of astrocyte-derived signals: implications for spinal cord injury

doi: 10.1186/s12974-020-01748-x

Figure Lengend Snippet: Astrocyte-derived CCL2 and CCL9 but not CCL1 regulate macrophage polarization, in vitro. a Macrophage cultures were exposed to ODN 2088-treated astrocyte CM (ODN 2088-CM), in the absence or presence of CCL1 neutralizing Ab. The graph shows the quantification of the F4/80 + /Arg-1 + cell number expressed as percentage of total F4/80 + cells in the macrophage cultures [ p = 0.7228, independent-sample t -test, two-tailed]. b Macrophage cultures were exposed to vehicle-treated astrocyte CM, in the absence or presence of CCL2 neutralizing Ab. The graph shows the quantification of the F4/80 + /Arg-1 + cell number expressed as percentage of total F4/80 + cells in the macrophage cultures [** p < 0.01, independent-sample t -test, two-tailed]. c Macrophage cultures were exposed to vehicle-treated astrocyte CM, in the absence or presence of CCL9 neutralizing Ab. The graph shows the quantification of the F4/80 + /Arg-1 + cell number expressed as percentage of total F4/80 + cells in the macrophage cultures [*** p < 0.001, independent-sample t -test, two-tailed]. d Macrophage cultures were exposed to vehicle-treated astrocyte CM (Veh-CM) or ODN 2088-treated astrocyte CM (ODN 2088-CM) for 24 h, with or without (control) addition of rmCCL9 (20 pg/ml). The graph shows the quantification of the F4/80 + /Arg-1 + double-labeled cells expressed as percent of total F4/80 + cells in macrophage cultures [ F (2, 6) = 53.68, p < 0.0001 by one-way ANOVA, * p < 0.05, ** p < 0.01, *** p < 0.001 by Tukey’s post hoc test]. The experiments were independently repeated twice, yielding similar results. Results from a representative experiment are shown. Results obtained from additional biological repeats of these experiments can be found in Additional file D-G. Data are presented as mean ± SEM

Article Snippet: In addition, CCL1 concentrations were independently measured by a second ELISA kit purchased from a different vendor (Rockland Immunochemicals; Limerick, PA, USA).

Techniques: Derivative Assay, In Vitro, Two Tailed Test, Control, Labeling

Journal: Journal of Neuroinflammation

Article Title: Astroglial TLR9 antagonism promotes chemotaxis and alternative activation of macrophages via modulation of astrocyte-derived signals: implications for spinal cord injury

doi: 10.1186/s12974-020-01748-x

Figure Lengend Snippet: A scheme summarizing the effects of ODN 2088-treated astrocytes on macrophages. TLR9 antagonism increases the release of CCL1 by astrocytes, which enhances macrophage chemotaxis. In contrast, CCL2 and CCL9 release are decreased in response to ODN 2088. This reduces the negative regulatory effect of CCL2 and CCL9 on M2 macrophage polarization and fosters the M2 phenotype

Article Snippet: In addition, CCL1 concentrations were independently measured by a second ELISA kit purchased from a different vendor (Rockland Immunochemicals; Limerick, PA, USA).

Techniques: Chemotaxis Assay

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

Journal: Frontiers in Immunology

Article Title: The diagnostic accuracy of CC chemokine ligand 23 for Kawasaki disease

doi: 10.3389/fimmu.2026.1758367

Figure Lengend Snippet: Intergroup comparison of CCL23 protein expression among the Kawasaki disease (KD) group, fever control (FC) group, and healthy control (HC) group.

Article Snippet: Meanwhile, the quantitative detection of CCL23 protein was conducted using the Human CCL23 ELISA Kit (Boster Biological Technology, Product No.: EK1224) in strict accordance with the kit instructions.

Techniques: Comparison, Expressing, Control

Journal: Frontiers in Immunology

Article Title: The diagnostic accuracy of CC chemokine ligand 23 for Kawasaki disease

doi: 10.3389/fimmu.2026.1758367

Figure Lengend Snippet: Diagnostic accuracy analysis of CC Chemokine Ligand 23 (CCL23) for Kawasaki disease (KD). (A) Comparison between the KD group and the fever control (FC) group; (B) Comparison between the KD group and the healthy control (HC) group; (C) Comparison between the KD group and the combined FC and HC groups.

Article Snippet: Meanwhile, the quantitative detection of CCL23 protein was conducted using the Human CCL23 ELISA Kit (Boster Biological Technology, Product No.: EK1224) in strict accordance with the kit instructions.

Techniques: Diagnostic Assay, Comparison, Control