Journal: Viruses

Article Title: Aerosolized Exposure to H5N1 Influenza Virus Causes Less Severe Disease Than Infection via Combined Intrabronchial, Oral, and Nasal Inoculation in Cynomolgus Macaques

doi: 10.3390/v13020345

Figure Lengend Snippet: Cytokine and chemokine expression levels in serum. Shown are IL-6, CXCL10, CCL2, CXCL11, CCL3 and CCL4 in pg/mL in serum, for each individual animal in time for the animals that received combined-route virus inoculation (left side of each graph), aerosol delivery (middle of each graph), or aerosol delivery with BAL collection (right side of each graph). Individual animal numbers are the same for all graphs and only indicated in the upper left graph.

Article Snippet: Cytokine and chemokine concentrations, including interleukin (IL-1β), IL-6, CCL11 (Eotaxin), CXCL10 (IP-10), CXCL11 (I-TAC), CCL2 (MCP-1), CXCL9 (MIG), CCL3 (MIP-1𝛼), CCL4 (MIP-1β), CCL5 (RANTES), CXCL8 (IL-8), TNF𝛼, and IFNγ, were determined using a LEGENDplexTM NHP Chemokine/Cytokine Panel (13-plex) (Biolegend, San Diego, CA, USA) according to the manufacturer’s instruction.

Techniques: Expressing, Virus, Aerosol

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. Cxcl9, Cxcl10 and Cxcl11 transcript levels were measured in islets isolated from 10 week old male (n = 4) and female NOD mice (n = 5). Data are expressed relative to age-matched BALB/c control mice (n = 7). **p<0.01, *p<0.05. B–F. Islets from Wistar rats (B, C; n = 4 per group) or human islets (D–F; n =3 per group) were untreated (NT) or stimulated with 10 ng/mL IL-1β, 100 U/mL IFN-γ or both cytokines for 3 h. B–F. Relative mRNA abundance of CXCL9 (B, D), CXCL10 (E) and CXCL11 (C, F) was determined by RT-PCR. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Isolation, Control, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A–C. 832/13 rat insulinoma cells were stimulated with 1 ng/mL IL-1β or IL-1β plus 100 U/mL IFN-γ for the indicated times (NT; no treatment). D–F. 832/13 cells were pretreated for 1 h with either DMSO or 0.5 μg/mL Cycloheximide (CHX). Cells were subsequently exposed to IL-1β (1 ng/mL) or the combination of IL-1β and IFN-γ (100 U/mL) for 2 h. Cellular mRNA levels of Cxcl9 (A, D), Cxcl10 (B, E) and Cxcl11 (C, F) were detected by RT-PCR. n.s. = not significant vs respective treatment in DMSO control group. Data are shown as means ± SEM from three independent experiments.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were exposed to 100 U/mL IFN-γ for the indicated times. PO4-STAT1Y701 and total STAT1 protein abundance were determined by immunoblotting. B, C. 832/13 cells were pre-treated for 1 h with increasing concentrations of JAKi (1 nM, 10 nM, 100 nM), followed by a 3 h stimulation with IL-1β alone (1 ng/mL) or IL-1β plus 100 U/mL IFN-γ. ***p<0.001 vs. DMSO (black bar), *p<0.05 vs. DMSO (black bar). D, E. 832/13 cells were transfected with two siRNA duplexes targeting STAT1 using a scrambled siRNA sequence duplex as a control. 48 h post- transfection cells were cultured for 3 h with 1 ng/ml IL-1β or IL-1β plus 100 U/ml IFN-γ. ***p<0.001 vs. siScramble (black bar), *p<0.05 vs. siScramble (black bar). Cxcl9 (B, D) and Cxcl11 (C, E) mRNA levels were quantified. Data are represented as means ± SEM from three independent experiments. The immunoblot in A was repeated on two separate occasions.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Quantitative Proteomics, Western Blot, Transfection, Sequencing, Control, Cell Culture

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: A. 832/13 cells were transduced with adenoviruses encoding either βGAL, wild-type STAT1 (WT), STAT1Y701F, STAT1S727A, STAT1Y701F/S727A (DM; double mutant) or STAT1S727T. STAT1 abundance was determined by immunoblotting. B, C. 832/13 cells were transduced with the adenoviruses indicated in (A); 24 h post-transduction cells were stimulated for 3 h with either IL-1β (1 ng/mL) alone or IL-1β plus IFN-γ (100 U/mL). *p<0.05, #p<0.1. D, E. Rat islets were transduced with the indicated adenoviruses. 24 h post-transduction cells were stimulated with both IL-1β (10 ng/mL) and IFN-γ (100 U/mL) for 3 h. ***p<0.001, **p<0.01. Relative mRNA abundance of Cxcl9 (B, D) and Cxcl11 (C, E) was determined by RT-PCR. Date are expressed as means ± SEM from 3 (B, C) or 2 (D, E) individual experiments. The immunoblot in A was repeated on two individual occasions.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques: Transduction, Mutagenesis, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: BioFactors (Oxford, England)

Article Title: Pancreatic β-cell Production of CXCR3 Ligands Precedes Diabetes Onset

doi: 10.1002/biof.1304

Figure Lengend Snippet: Schematic representation of distal and proximal GAS sites in the Cxcl9 and Cxcl11 promoters are shown (left panels). Arrows are the diagrammatic depiction of PCR amplicons. A–D. 832/13 cells were stimulated with 100 U/mL IFN-γ for either 20 mins (middle panels) or a time course (right panels). ChIP assays were performed to determine relative occupancy of total STAT1 (middle panels) and PO4-STAT1Y701 (right panels) on the Cxcl9 proximal (A) and distal promoter (B), and on the Cxcl11 proximal (C) and distal (D) promoter. ***p<0.001 vs. NT, **p<0.01 vs. NT, *p<0.05 vs. NT. Data are expressed as means ± SEM from 3–4 individual experiments.

Article Snippet: Serum levels of CXCL9, CXCL10, and CXCL11 were detected using the mouse CXCL9 Quantikine ELISA kit (Cat # MCX900), CXCL10 Quantikine ELISA kit (Cat # MCX100) and the mouse CXCL11 DuoSet ELISA kit (Cat # DY572), all from R&D Systems (Minneapolis, MN) using the protocol provided by the manufacturer.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Patient Derived Colonoids as Drug Testing Platforms–Critical Importance of Oxygen Concentration

doi: 10.3389/fphar.2021.679741

Figure Lengend Snippet: Chemokines released by colonoids at 20 or 2% O 2 . Conditioned medium from colonoids at 20 or 2% O 2 , followed by 24h treatment with TNF, IL17 or TNF/IL17, and untreated colonoids were analyzed for chemokine concentration (A) Mean protein concentration ( p g . ml −1 ) of chemokines in conditioned medium from colonoids ( n = 3 independent assays) analyzed by Human Chemokine Panel featuring 40 magnetic bead-based immunoassays. The 18 heatmaps show expression of significantly regulated proteins and are grouped according to 1) Highest protein expression by TNF/IL17 treatment at 20% O 2 . 2) Highest protein expression by TNF/IL17 treatment at 2% O 2 . 3) Highest protein expression by TNF treatment at 20% O 2 , and 4) Highest protein expression by TNF or TNF/IL17 treatment, similar at 20 and 2% O 2 (B) CCL20, CXCL1, CXCL2, CXCL5, CXCL8, CXCL10 and CXCL11 protein concentrations ( p g.ml −1 ) in conditioned medium from minimum six assays measured by ELISA. Left panels: paired data at 2 or 20% O 2 across all treatments for the selected chemokines, analyzed with Wilcoxon matched-pairs signed rank test. Significance level is indicated as p values or not significant (ns). Right panels: concentrations for each chemokine in 2% (blue) or 20% (red) O 2 plotted as individual values with mean and SD. Statistical analysis were performed on log2 transformed data . Alterations across different treatments within each oxygen concentrations were analyzed with one-way ANOVA followed by Tukey’s multiple comparisons or Kruskal-Wallis test followed by Dunn’s multiple comparisons test. Alterations across different oxygen concentrations within same treatment were analyzed by two-way ANOVA followed by Holm-Šídák multiple comparisons test. * p < 0.05; red and blue asterisks show comparisons across different treatments within 20 and 2% O 2 conditions, respectively. Black asterisks indicate significant comparisons between 2 and 20% oxygen concentrations within a specific treatment.

Article Snippet: ELISA kits for human CXCL1 (DY275-05), CXCL2 (DY276-05), CXCL5 (DY254-05), CXCL8 (DY208), CXCL10 (DY266-05), CXCL11 (DY672) and CCL20 (DY360), all from R&D Systems (Abington, United Kingdom), were used according to manufacturer’s protocols.

Techniques: Concentration Assay, Protein Concentration, Expressing, Enzyme-linked Immunosorbent Assay, Transformation Assay

Journal: Journal of Translational Medicine

Article Title: In vitro migration of cytotoxic T lymphocyte derived from a colon carcinoma patient is dependent on CCL2 and CCR2

doi: 10.1186/1479-5876-9-33

Figure Lengend Snippet: Chemokine receptors expressed by CTL007, and chemokines produced by WC007 CRC cells

Article Snippet: Recombinant human CXCL11 was purchased from R&D Systems.

Techniques: Produced, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Pharmacology

Article Title: CXCR7 Antagonism Reduces Acute Lung Injury Pathogenesis

doi: 10.3389/fphar.2021.748740

Figure Lengend Snippet: Kinetics of the expression of CXCR3/CXCR4 and its ligands in the BAL of LPS-challenged DBA/1 mice. ALI was induced by nebulized LPS inhalation in male DBA/1 mice. Control mice inhaled NaCl 0.9% ( n = 12 mice; all time points were pooled). (A) LPS inhalation increased protein concentrations of the CXCR3 ligands CXCL9, CXCL10, and CXCL11, measured in the BAL 5, 24, 48, and 72 h following LPS challenge, compared with control mice. Results are expressed as mean ± SEM ( n = 8 mice per time point). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus control mice using Student t -test. (B) Representative gating strategy for CXCR3 + myeloid and lymphoid cells. (C) CXCR3 expression on BAL lymphoid and myeloid-infiltrating cells. Results are expressed as mean ± SEM of the mean fluorescence intensity (MFI) of CXCR3 obtained for each LPS-challenged mouse ( n = 7–8 mice per time point) corrected by the MFI obtained in the fluorescence minus one (FMO) controls for CXCR3. Negative MFI values were set to 0. The complete gating strategy for lymphoid and myeloid cells is illustrated in . (D) Proportion of CXCR3 + lymphoid and myeloid cells expressed as percentages (mean ± SEM) of the CD45 + CD11b − and CD45 + CD11b + parent population, respectively, in the BAL (see gating strategy in ) ( n = 7–8 mice per time point). (E) Time course of BAL CXCR3 + lymphoid and myeloid cell infiltrates. Results are expressed as absolute counts in the BAL (mean ± SEM). ** p < 0.01, *** p < 0.001, **** p < 0.0001 using Student t -test versus control mice. (F) LPS inhalation increased protein concentrations of the CXCR4 ligand CXCL12, measured in the BAL 5, 24, 48, and 72 h following LPS nebulization compared with control mice. Results are expressed as mean ± SEM ( n = 7–8 mice per time point). ** p < 0.01, **** p < 0.0001 versus control mice using Student t -test. (G) Representative gating strategy for CXCR4 + myeloid and lymphoid cells. (H) CXCR4 expression on BAL lymphoid and myeloid-infiltrating cells. Results are expressed as mean ± SEM of the MFI of CXCR4 obtained for each LPS-challenged mouse ( n = 7–8 mice per time point) corrected by the MFI obtained in the FMO controls for CXCR4. Negative MFI values were set to 0. The gating strategy for lymphoid and myeloid cells is illustrated in . (I) Proportion of CXCR4 + lymphoid and myeloid cells expressed as percentages (mean ± SEM) of the CD45 + CD11b − and CD45 + CD11b + parent population, respectively, in the BAL (see gating strategy in ) ( n = 7–8 mice per time point). (H) Time course of BAL CXCR4 + lymphoid and myeloid cell infiltrates. Results are expressed as absolute counts in the BAL (mean ± SEM) ( n = 7–8 mice per time point). ** p < 0.01, *** p < 0.001, **** p < 0.0001 using Student t -test versus control mice.

Article Snippet: Recombinant murine CXCL11 (250-29; Peprotech, Cranbury, NJ, United States) was used as a standard, and Fluor-labeled anti-mouse CXCL11 polyclonal antibody (AF572; R&D Systems) was used as the detection antibody.

Techniques: Expressing, Control, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: CXCR7 Antagonism Reduces Acute Lung Injury Pathogenesis

doi: 10.3389/fphar.2021.748740

Figure Lengend Snippet: Antagonism of CXCR7 increases plasma CXCL11 and CXCL12 levels and decreases CXCR3 + and CXCR4 + BAL infiltrates post LPS challenge. ALI was induced by nebulized LPS inhalation, and DBA/1 mice were treated with vehicle (LPS-vehicle, black bars) or ACT-1004-1239 (LPS-ACT-1004-1239, 100 mg/kg, red bars) orally, twice daily, 1 h prior to LPS challenge. Control mice received vehicle 1 h prior to NaCl 0.9% inhalation (NaCl-vehicle, white bars; n = 12 mice, pool of all time points). Protein concentrations of CXCL11 and CXCL12 in the plasma and lung tissue and BAL flow cytometry of immune infiltrates were performed 24, 48, and 72 h after challenge ( n = 6–8 mice per time point). Time course of CXCL11 protein concentration in the plasma (A) and lung tissue (B) . Chemokine concentrations are expressed in pg/ml (plasma) or pg/lung homogenate (mean ± SEM). ** p < 0.01, *** p < 0.001 versus LPS-vehicle-treated animals or # p < 0.05, ### p < 0.001 versus NaCl-vehicle-treated control mice using Student t -tests. Time course of BAL CXCR3 + lymphoid (C) and myeloid (D) infiltrates. Results are expressed as absolute cell counts in the BAL (mean ± SEM). * p < 0.05, **** p < 0.0001 versus LPS-vehicle-treated animals or ### p < 0.001, #### p < 0.0001 versus NaCl-vehicle-treated control mice using Student t-tests. Time course of CXCL12 concentration in the plasma (E) and lung tissue (F) . Chemokine concentrations are expressed in ng/ml (plasma) or ng/lung homogenate (mean ± SEM). **** p < 0.0001 versus LPS-vehicle-treated animals or # p < 0.05, ### p < 0.001 versus NaCl-vehicle-treated control mice using Student t-tests. Time course of BAL CXCR4 + lymphoid (G) and BAL CXCR4 + myeloid (H) infiltrates. Results are expressed as absolute counts in the BAL (mean ± SEM). * p < 0.05, ** p < 0.01, using Student t-test versus LPS-vehicle-treated animals or ### p < 0.001, #### p < 0.0001 versus NaCl-vehicle-treated control mice using Student t-tests.

Article Snippet: Recombinant murine CXCL11 (250-29; Peprotech, Cranbury, NJ, United States) was used as a standard, and Fluor-labeled anti-mouse CXCL11 polyclonal antibody (AF572; R&D Systems) was used as the detection antibody.

Techniques: Clinical Proteomics, Control, Flow Cytometry, Protein Concentration, Concentration Assay

Journal: Frontiers in Pharmacology

Article Title: CXCR7 Antagonism Reduces Acute Lung Injury Pathogenesis

doi: 10.3389/fphar.2021.748740

Figure Lengend Snippet: Treatment with ACT-1004-1239 dose-dependently increases plasma CXCL11 and CXCL12 levels and reduces BAL T cell and inflammatory macrophage infiltrates in the LPS-induced ALI/ARDS model. Vehicle (Veh; black bars) or ACT-1004-1239 (10, 30, or 100 mg/kg; bars with different shades of red) was given orally, twice daily, starting 1 h prior to LPS nebulization, for a total of six administrations. Control mice (white bars) were challenged by NaCl nebulization and received vehicle administrations. Plasma CXCL11 (A) and plasma CXCL12 levels (B) 72 h after LPS or NaCl challenge. Results are expressed as mean + SEM ( n = 10–25 mice per treatment-LPS groups and n = 4–5 mice for control group). * p < 0.05, **** p < 0.0001 versus vehicle-treated LPS-challenged mice, using one-way ANOVA test followed by Dunnett’s multiple comparisons test. Total BAL T cell (C) and BAL inflammatory macrophage counts (D) 72 h after LPS challenge. Results are expressed as mean ± SEM with n = 11–23 mice per treatment-LPS groups and n = 3 for controls. ** p < 0.01, *** p < 0.001, **** p < 0.0001 versus vehicle-treated LPS-challenged mice, using one-way ANOVA test followed by Dunnett’s multiple comparisons test.

Article Snippet: Recombinant murine CXCL11 (250-29; Peprotech, Cranbury, NJ, United States) was used as a standard, and Fluor-labeled anti-mouse CXCL11 polyclonal antibody (AF572; R&D Systems) was used as the detection antibody.

Techniques: Clinical Proteomics, Control

Journal: Frontiers in Pharmacology

Article Title: CXCR7 Antagonism Reduces Acute Lung Injury Pathogenesis

doi: 10.3389/fphar.2021.748740

Figure Lengend Snippet: Summary of pathological role of CXCR7 and potential benefit of CXCR7 antagonism during ALI/ARDS. (A) CXCR7 functions predominantly as a scavenger receptor for its two ligands: the interferon-inducible chemokine CXCL11 and the constitutive chemokine CXCL12. Binding of its ligands leads to internalization of the CXCR7–ligand complex and ligand degradation. CXCL11 and CXCL12 also bind and activate the signaling chemokine receptors CXCR3 and CXCR4, respectively. The CXCR3/CXCR4/CXCR7 axes play an important role in lung inflammation. CXCR7 scavenging activity tightly regulates the extracellular levels of its ligands, facilitating the establishment and maintenance of CXCL11/12 chemokine concentration gradients and CXCR3 + /CXCR4 + cell migration from the blood to the inflamed lung. In addition, increased CXCR7 expression in the inflamed lung has been reported to be associated with breathing pattern alteration and endothelial barrier dysfunction. (B) CXCR7 antagonism with the CXCR7 antagonist ACT-1004-1239 exhibits immunomodulatory effects: by blocking the scavenging activity of the receptor and consequently increasing CXCL11 and CXCL12 plasma concentrations, chemokine gradients are disrupted, inhibiting CXCR3 + and CXCR4 + cell migration to the inflamed lung. In addition, treatment with ACT-1004-1239, by increasing plasma CXCL12 and/or by direct inhibition of CXCR7 signaling, ameliorates ALI-induced breathing pattern alteration and endothelial barrier dysfunction.

Article Snippet: Recombinant murine CXCL11 (250-29; Peprotech, Cranbury, NJ, United States) was used as a standard, and Fluor-labeled anti-mouse CXCL11 polyclonal antibody (AF572; R&D Systems) was used as the detection antibody.

Techniques: Binding Assay, Activity Assay, Concentration Assay, Migration, Expressing, Blocking Assay, Clinical Proteomics, Inhibition

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The expression and possible roles of chemokine CXCL11 and its receptor CXCR3 in the human endometrium.

doi: 10.4049/jimmunol.177.12.8813

Figure Lengend Snippet: FIGURE 2. IFN--stimulated CXCL11 protein production in EEC and ESC. A, EEC and ESC were cultured in serum-free medium with different doses of IFN- for 24 h. Cell extracts were prepared and assayed for CXCL11 by Western blotting. The result is representative of three separate experiments. B, EEC and ESC were cultured in serum-free medium with different doses of IFN- for 24 h. The conditioned medium were collected and assayed for CXCL11 concentrations by ELISA. The values were nor- malized with total protein of cell extracts. Values are the mean SEM of the combined data of five separate experiments using different EEC and ESC preparations. #, p 0.0001, between EEC and ESC with IFN- at 10, 100, 1000 ng/ml. , p 0.01; , p 0.0001, both vs control of EEC; , p 0.0005, both vs control of ESC.

Article Snippet: E-mail address: yutakaos-tky@umin.ac.jp Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 by guest on M arch 3, 2015 http://w w w .jim m unol.org/ D ow nloaded from recombinant IFN- , human recombinant CXCL11, and human recombinant IL-2 were obtained from R&D Systems.

Techniques: Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The expression and possible roles of chemokine CXCL11 and its receptor CXCR3 in the human endometrium.

doi: 10.4049/jimmunol.177.12.8813

Figure Lengend Snippet: FIGURE 3. IFN--induced protein expression of three CXCR3 ligands CXCL9, CXCL10, and CXCL11 in EEC. EEC and ESC were cultured in serum-free medium with different doses of IFN- for 24 h. Cell extracts were prepared and assayed for CXCL9, CXCL10, and CXCL11 by West- ern blotting. All the chemokines were expressed in dose-dependent man- ners. The result is representative of three separate experiments.

Article Snippet: E-mail address: yutakaos-tky@umin.ac.jp Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 by guest on M arch 3, 2015 http://w w w .jim m unol.org/ D ow nloaded from recombinant IFN- , human recombinant CXCL11, and human recombinant IL-2 were obtained from R&D Systems.

Techniques: Expressing, Cell Culture

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The expression and possible roles of chemokine CXCL11 and its receptor CXCR3 in the human endometrium.

doi: 10.4049/jimmunol.177.12.8813

Figure Lengend Snippet: FIGURE 5. Effects of conditioned medium of IFN--stimulated EEC on the migration of T cells and trophoblast cells. Migration assay was per- formed to study whether the migration of T cells and trophoblast cells was affected by endometrial CXCL11 expression. Supernatants of EEC either stimulated or not by IFN- (100 ng/ml) for 24 h were preincubated for 1 h with 10 g/ml anti-CXCL9 Ab, anti-CXCL10 Ab, anti-CXCL11 Ab, or isotype control rabbit IgG, and plated on the lower chambers. Cells were plated on the upper wells of Transwell membranes containing 100 l of serum-free DMEM/F12. 5 106 T cells (A) were incubated for 2 h, and 2 105 trophoblast cells (B) were for 24 h. After the incubation, T cells and trophoblast cells on the upper surface of membranes were completely removed and migrated cells were fixed with acetone/methanol. Migration indices were determined by counting the cell number. The values represent relative ratios of the cell number compared with those in using the control supernatants of EEC with rabbit IgG. A, Values are the mean SEM of the combined data from three independent experiments using different T cell preparations. , p 0.05, control with rabbit IgG vs IFN- with rabbit IgG. , p 0.001, each vs IFN- with rabbit IgG. B, Values are the mean SEM of the combined data from three independent experiments using dif- ferent trophoblast cell preparations. , p 0.05, control plus rabbit IgG vs IFN- plus rabbit IgG. , p 0.05, each vs IFN- plus rabbit IgG.

Article Snippet: E-mail address: yutakaos-tky@umin.ac.jp Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 by guest on M arch 3, 2015 http://w w w .jim m unol.org/ D ow nloaded from recombinant IFN- , human recombinant CXCL11, and human recombinant IL-2 were obtained from R&D Systems.

Techniques: Migration, Expressing, Control, Incubation

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The expression and possible roles of chemokine CXCL11 and its receptor CXCR3 in the human endometrium.

doi: 10.4049/jimmunol.177.12.8813

Figure Lengend Snippet: FIGURE 6. CXCL11-induced ESC proliferation via p42/44 MAPK ac- tivation. A, The effect of CXCL11 on the proliferation of ESC was exam- ined by measuring BrdU incorporation into DNA by using the cell prolif- eration ELISA. ESC were treated with CXCL11 at different concentrations for 24 h. The values represent relative ratios compared with those in un- treated cells. Values are the mean SEM of the combined data from five independent experiments using different ESC preparations. , p 0.005; , p 0.0005, both vs control. B, ESC were incubated with 100 ng/ml CXCL11 for the indicated times (0–240 min). Cell extracts were prepared and assayed for phosphorylated p42/44 MAPK (phospho-p42/44) or total p42/44 MAPK (total-p42/44) by Western blotting. The result is represen- tative of three separate experiments. C, Effects of MEK inhibitor PD98059 on CXCL11-induced cell proliferation of ESC was examined by measuring BrdU incorporation into DNA by using the cell proliferation ELISA. ESC were treated with or without PD98059 (25 M), for 1 h, and then stimu- lated with CXCL11 (100 ng/ml). After 24 h incubation, BrdU incorpora- tion into DNA in ESC was measured using the cell proliferation ELISA. The values represent relative ratios compared with those in untreated cells. Values are the mean SEM of the combined data from four independent experiments using different ESC preparations. , p 0.0001, control vs CXCL11. , p 0.0001 CXCL11 vs CXCL11 with PD98059.

Article Snippet: E-mail address: yutakaos-tky@umin.ac.jp Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 by guest on M arch 3, 2015 http://w w w .jim m unol.org/ D ow nloaded from recombinant IFN- , human recombinant CXCL11, and human recombinant IL-2 were obtained from R&D Systems.

Techniques: BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay, Control, Incubation, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The expression and possible roles of chemokine CXCL11 and its receptor CXCR3 in the human endometrium.

doi: 10.4049/jimmunol.177.12.8813

Figure Lengend Snippet: FIGURE 7. CXCL11-induced inhibition of proliferation and stimulation of apoptosis in EEC. A, The effect of CXCL11 on the proliferation of EEC was examined by measuring BrdU incorporation into DNA by using the cell proliferation ELISA. EEC were treated with CXCL11 at different concentrations for 24 h. The values represent relative ratios compared with those in untreated cells. Values are the mean SEM of the combined data from four independent experiments using different EEC preparations. , p 0.05; , p 0.0001, both vs control. B, The effect of CXCL11 on cell death of EEC was determined by the measurement of LDH in CXCL11-treated EEC supernatant. EEC were treated with CXCL11 at 100 ng/ml or with IFN- at 100 ng/ml for 24 h. The values represent relative ratios compared with those in untreated cells. Values are the mean SEM of the combined data from three independent experiments using different EEC preparations. , p 0.05; , p 0.0001, both vs control. C, The effect of CXCL11 on apoptosis of EEC was determined by double staining of annexin V and PI. EEC were treated with CXCL11 at 100 ng/ml or with IFN- at 100 ng/ml for 48 h. The cells were stained with Annexin VFITC and PI. Apoptosis was analyzed by flow cytometry on 5 104 EEC. The result is representative of four separate experiments. D, The percentage of apoptotic EEC treated with CXCL11 and IFN- was significantly higher than that of the control. Annexin V-positive cells were regarded as apoptotic cells. Values are the mean SEM of the combined data from four independent experiments using different EEC preparations. , p 0.05, both vs control.

Article Snippet: E-mail address: yutakaos-tky@umin.ac.jp Copyright © 2006 by The American Association of Immunologists, Inc. 0022-1767/06/$02.00 by guest on M arch 3, 2015 http://w w w .jim m unol.org/ D ow nloaded from recombinant IFN- , human recombinant CXCL11, and human recombinant IL-2 were obtained from R&D Systems.

Techniques: Inhibition, BrdU Incorporation Assay, Enzyme-linked Immunosorbent Assay, Control, Double Staining, Staining, Cytometry

Journal: Free radical biology & medicine

Article Title: Oxidative stress-induced decreased expression of FABP5 leads to mitochondrial damage and survival disorder of decidual stromal cells in women with recurrent spontaneous abortion.

doi: 10.1016/j.freeradbiomed.2025.06.003

Figure Lengend Snippet: Fig. 7. FABP5 downregulation in DSCs repressed CXCL11/CXCR3 signaling between DSCs and HTR-8/Svneo cells. (A) KEGG pathway analysis of DEGs identified by RNA-seq in FABP5-knockdown primary DSC. (B) Gene Set Enrichment Analysis (GSEA) of chemokine signaling pathways in FABP5-silenced DSCs. (C) FPKM values of CXCL11 expression in control and FABP5-knockdown groups. (D) RT-qPCR, (E, F) Western blot, and (G) ELISA quantification of CXCL11 expression in DSCs transfected with FABP5-targeting or NC siRNAs. (H) RT-qPCR and (I, J) Western blot analyses of CXCR3 expression in HTR-8/Svneo cells treated with CS from FABP5-deficient or control DSCs. Statistical significance: *P < 0.05, **P < 0.01.

Article Snippet: The concentration of CXCL11 in the CS was measured using a human CXCL11 ELISA kit (Boster, Beijing, China; CAT# EK0737) following the manufacturer’s instructions.

Techniques: RNA Sequencing, Knockdown, Protein-Protein interactions, Expressing, Control, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection