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: 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

Journal: The Journal of Cell Biology

Article Title: Regulation of C-X-C chemokine gene expression by keratin 17 and hnRNP K in skin tumor keratinocytes

doi: 10.1083/jcb.201408026

Figure Lengend Snippet: CXCR3 inhibition results in an attenuated transformed phenotype. (A) CXCL11 protein amounts in conditioned medium from A431 SCR or KRT17 shRNA cells were measured by ELISA. Data from three experimental repeats (each performed in 10 replicates) are represented as mean ± SEM (error bars). *, P < 0.016. (B) CXCL9 protein amounts in conditioned medium from primary keratinocytes isolated from ear lesions of P81 Krt17 +/+ or Krt17 −/− Gli2 tg/+ mice were measured by ELISA. Data from four experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.04. (C) CXCL11 protein amounts in conditioned medium from A431 SCR shRNA cells transfected with either NS or hnRNP K siRNA (RPK1) were measured by ELISA. Data from three experimental repeats (each performed in 10 replicates) are represented as mean ± SEM (error bars). *, P < 0.04. (D) Immunoblotting performed on lysates from A431 cells stably expressing SCR or KRT17 shRNA with antibodies against the indicated proteins. (E) Soft agar colony assay using A431 SCR shRNA cells. Cells were grown in conditioned medium from A431 SCR or KRT17 shRNA cells, supplemented with anti-CXCR3 neutralizing antibody (0.5 ng/ml) or IgG control. Bar, 1 cm. (F) Colony numbers from E were quantitated using ImageJ and normalized to those grown in IgG-supplemented conditioned medium from A431 SCR shRNA cells. Data from four experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.03. (G) Soft agar colony assay using A431 SCR shRNA cells. Cells were grown in conditioned medium from A431 SCR shRNA cells, supplemented with DMSO or CXCR3 antagonist (10 µM). Colony numbers were quantitated using ImageJ and normalized to those grown in DMSO control. Data from three experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.05. (H) Soft agar colony assay using A431 SCR shRNA cells. Cells were grown in conditioned medium from A431 KRT17 shRNA cells, supplemented with or without 10 ng/ml recombinant CXCL11. Bar, 1 cm. (I) Colony numbers from H were quantitated using ImageJ and normalized to those grown without CXCL11. Data from five experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.02. (J) Soft agar colony assay using A431 SCR shRNA cells. Cells were grown in conditioned medium from A431 cells transfected with mCherry-C1 vector or hnRNP K, supplemented with anti-CXCR3 neutralizing antibody (0.5 ng/ml) or IgG control. Colony numbers were quantitated using ImageJ and normalized to those grown in conditioned medium from vector-expressing cells. Data from four experimental repeats (each performed in triplicate) are represented as mean ± SEM (error bars). *, P < 0.02; **, P < 0.05.

Article Snippet: After 48 h, the medium was collected and CXCL9, CXCL11, CXCL8, IL-17F, and TSLP levels were determined using mouse CXCL9/MIG Duoset, human CXCL11/I-TAC Duoset, human CXCL8/IL-8 Duoset, human IL-17F Duoset, and human TSLP Duoset (all from R&D Systems) according to the manufacturer’s protocol.

Techniques: Inhibition, Transformation Assay, shRNA, Enzyme-linked Immunosorbent Assay, Isolation, Transfection, Western Blot, Stable Transfection, Expressing, Colony Assay, Control, Recombinant, Plasmid Preparation

Journal: The Journal of Cell Biology

Article Title: Regulation of C-X-C chemokine gene expression by keratin 17 and hnRNP K in skin tumor keratinocytes

doi: 10.1083/jcb.201408026

Figure Lengend Snippet: A model depicting the significance of K17- and hnRNP K–dependent expression of CXCR3 ligands. K17 binds to hnRNP K and is required for its cytoplasmic localization. hnRNP K binds to and regulates mRNA transcripts for the CXCR3 ligands CXCL9, CXCL10, and CXCL11 in a K17-dependent manner. The K17–hnRNP K interaction and expression of CXCR3 ligands are both regulated by RSK activity. CXCR3 ligand secretion and CXCR3 signaling are required for K17- and hnRNP K–dependent enhancement of tumor cell growth and invasion.

Article Snippet: After 48 h, the medium was collected and CXCL9, CXCL11, CXCL8, IL-17F, and TSLP levels were determined using mouse CXCL9/MIG Duoset, human CXCL11/I-TAC Duoset, human CXCL8/IL-8 Duoset, human IL-17F Duoset, and human TSLP Duoset (all from R&D Systems) according to the manufacturer’s protocol.

Techniques: Expressing, Activity Assay

Journal: The Journal of Cell Biology

Article Title: Regulation of C-X-C chemokine gene expression by keratin 17 and hnRNP K in skin tumor keratinocytes

doi: 10.1083/jcb.201408026

Figure Lengend Snippet: Oligonucleotide primer sets used for qRT-PCR assays

Article Snippet: After 48 h, the medium was collected and CXCL9, CXCL11, CXCL8, IL-17F, and TSLP levels were determined using mouse CXCL9/MIG Duoset, human CXCL11/I-TAC Duoset, human CXCL8/IL-8 Duoset, human IL-17F Duoset, and human TSLP Duoset (all from R&D Systems) according to the manufacturer’s protocol.

Techniques:

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