Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: LM2 Cells Transduced with Lenti-mCXCL9 Have Reduced Tumorigenicity In Vivo Compared with LM2 (A) Concentration of mCXCL9 levels in the supernatants of LM2 cells transduced with Lenti-mCXCL9. ELISA data are shown at 24 h after plating in triplicate + standard deviation (****p < 0.0001). (B) Viability of LM2-Lenti-mCXCL9 compared with LM2 cells in vitro . Data are shown as mean percent cell viability in comparison with mock-infected cells at 24, 48, and 72 h postinfection + standard deviation. (C) Survival of mice bearing LM2 or Lenti-transduced LM2 tumors was evaluated using Kaplan-Meier survival curves (*p < 0.05).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: Transduction, In Vivo, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, In Vitro, Infection

Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: Murine and Human CXCL9 Transgenes Engineered in Recombinant VSVs Do Not Alter the Growth Kinetics or Viral Killing Ability In Vitro Schematic depiction of the genomes of recombinant VSVs encoding murine (A) or human (B) CXCL9, CXCLi, and GFP. (C) Replication kinetics of VSVs encoding murine or human CXCL9 were performed in a multistep viral growth curve in Vero cells. Data are shown from duplicate experiments as average titer + standard deviation. (D) Viability of VSV-mCXCL9-, VSV-mCXCLi-, and VSV-GFP-infected Vero and murine LM2 tumor cells was assessed at 24 and 48 h postinfection at an MOI of 10. (E) Viability of VSV-M51R-hCXCL9- and VSV-M51R-hCXCLi-infected Vero and human FaDu-Luc tumor cells was measured at 24 and 72 h postinfection at an MOI of 10. Data are presented as duplicate experiments as average percent viability compared with mock-infected cells + standard deviation. Significance was determined by paired t test (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: Recombinant, In Vitro, Standard Deviation, Infection

Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: CXCL9 Expressed from VSV-mCXCL9 Is Biologically Active (A) Murine CXCL9 secretion was evaluated in vitro in the LM2 non-small cell lung cancer cell line. Supernatants of VSV-infected LM2 cells (MOI 0.1) were collected 24 h postinfection, and chemokine concentration was determined by ELISA. Concentrations are presented as average concentration + standard deviation. (B) Chemotactic activity of virally encoded mCXCL9 was assessed in an in vitro migration assay adapted from Campanella et al. Numbers of migrated cells are presented as average percent increase in migration compared with mock treated + standard deviation. Significance was determined by paired two-tailed t test (**p < 0.01).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: In Vitro, Infection, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation, Activity Assay, Migration, Two Tailed Test

Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: Oncolytic Activity of VSV-mCXCL9 Survival of mice bearing 5TGM1 or LM2 tumors was evaluated using Kaplan-Meier survival curves. (A) 5 × 10 6 5TGM1 cells were implanted subcutaneously on the right flanks of C57Bl6/KaLwRij mice; once tumors had grown to ∼6-mm diameter, mice were injected intravenously with PBS, VSV-mCXCL9, or VSV-GFP (n = 10 for PBS and VSV-mCXCL9 treatments, n = 9 for VSV-GFP treatment). (B) 1 × 10 6 LM2 cells were implanted subcutaneously on the right flanks of A/J mice, and established tumors (∼6-mm diameter) were injected intratumorally with PBS, VSV-mCXCL9, or VSV-GFP (n = 10 for all treatment groups). Survival analysis was performed using log rank statistics (***p < 0.001, ****p < 0.0001).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: Activity Assay, Injection

Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: Generation of a Tumor to Blood CXCL9 Chemokine Gradient after Intratumoral VSV-mCXCL9 Administration Intratumoral and serum mCXCL9 protein concentrations were determined after intratumoral injection of VSV-mCXCL9 or VSV-GFP in LM2 tumor-bearing mice by ELISA (n = 3 mice/group for each time point). Values are presented as average chemokine concentration in pg/mL + standard deviation. Significance was determined by paired two-tailed t test (**p < 0.01).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Standard Deviation, Two Tailed Test

Journal: Molecular Therapy Oncolytics

Article Title: Generation of a Tumor-Specific Chemokine Gradient Using Oncolytic Vesicular Stomatitis Virus Encoding CXCL9

doi: 10.1016/j.omto.2019.12.003

Figure Lengend Snippet: VSV-mCXCL9 Does Not Increase CD8 + or Th1 Cell Intratumoral Infiltration or CD8 + Inflammatory Cytokine Production LM2 tumors were harvested and processed 7 and 10 days after intratumoral (i.t.) injection of VSV-mCXCL9, VSV-GFP, or PBS, and flow data were gated on (A) CD45 + CD3 + CD8 + T cells or (C) CD45 + CD3 + CD4 + T-bet + (Th1) cells. T cell numbers are presented as average number of immune cells/mg tumor ± standard deviation (n = 4 mice/group/day). (B) Cytokine production was gated on TNF-α- and IFN-γ-producing cells from (A). Statistical significance was determined by paired two-tailed t test (*p < 0.05).

Article Snippet: Mouse cxcl9 was PCR amplified from a mCXCL9 cloning plasmid (catalog number MG50155-M; Sino Biological, Wayne, PA, USA).

Techniques: Injection, Standard Deviation, Two Tailed Test

Journal: Journal of Investigative Dermatology

Article Title: Differential Production of Th1- and Th2-Type Chemokines by Mouse Langerhans Cells and Splenic Dendritic Cells

doi: 10.1111/j.0022-202x.2004.23607.x

Figure Lengend Snippet: Figure 1 RT-PCR analysis of CXCL10, CXCL9, and CXCL11 gene expression in freshly isolated Langerhans cells (fLC), 48 h cultured LC (cLC), interferon-c (IFN-c)-stimulated cLC (c-cLC), freshly isolated splenic dendritic cells (fDC), 48 h cultured splenic DC (cDC), and IFN-c- stimulated cDC (c-cDC). LC and splenic DC were purified and cul- tured for 48 h in the absence or presence of 100 ng per mL of IFN-g. Samples from fresh and cultured LC and splenic DC were collected and mRNA expression was analyzed using specific primers for each chemokine. CXCL10, CXCL9, and CXCL11 mRNA expression was hardly detectable in fLC. In cLC, mRNA expression of CXCL10 and CXCL11, but not CXCL9, was induced. CXCL9 mRNA was strongly expressed only in g-cLC. In addition, mRNA for CXCL10 and CXCL11 was also strongly expressed in g-cLC. In splenic DC, mRNA for these T helper 1(Th1)-type chemokines was almost undetectable both in fDC and cDC. When stimulated with IFN-g, CXCL10, CXCL9, and CXCL11 mRNA expression was induced. Data are representative of three inde- pendent experiments.

Article Snippet: Measurement of CXCL9, CXCL11, and CCL17 Culture supernatants were collected, stored at 201C, and subjected to the quantification of protein levels of CXCL9, CXCL11, and CCL17 by ELISA using commercially available mouse CXCL9, CXCL11, and CCL17 immunoassay kits (R&D Systems), respectively, according to the manufacturer’s instructions.

Techniques: Reverse Transcription Polymerase Chain Reaction, Gene Expression, Isolation, Cell Culture, Expressing

Journal: Journal of Investigative Dermatology

Article Title: Differential Production of Th1- and Th2-Type Chemokines by Mouse Langerhans Cells and Splenic Dendritic Cells

doi: 10.1111/j.0022-202x.2004.23607.x

Figure Lengend Snippet: Figure 2 Production of T helper 1-type chemokines during culture of Langerhans cells (LC) and splenic dendritic cells (DC). Purified LC and splenic DC were cultured with or without interferon-g (IFN-g), and the concentration of CXCL10, CXCL9, and CXCL11 was measured at different time points (0, 12, 24, 36, 48 h) in the supernatants by ELISA. Representative data of three independent experiments.

Article Snippet: Measurement of CXCL9, CXCL11, and CCL17 Culture supernatants were collected, stored at 201C, and subjected to the quantification of protein levels of CXCL9, CXCL11, and CCL17 by ELISA using commercially available mouse CXCL9, CXCL11, and CCL17 immunoassay kits (R&D Systems), respectively, according to the manufacturer’s instructions.

Techniques: Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of Investigative Dermatology

Article Title: Differential Production of Th1- and Th2-Type Chemokines by Mouse Langerhans Cells and Splenic Dendritic Cells

doi: 10.1111/j.0022-202x.2004.23607.x

Figure Lengend Snippet: Figure 3 Chemotaxis of mCXCR3-transfected 2B4 T cells. Langerhans cells (LC) and splenic dendritic cells (DC) were cultured for 48 h with 100 ng per mL of interferon-g (IFN-g) and the supernatants were collected. The culture supernatants were preincubated with or without neutralizing anti-chemokine monoclonal antibody (mAb) indicated in the figure for 30 min, and assessed for chemotactic activity to CXCR3 transfectant. RPMI 10 medium alone served as a negative control. RPMI 10 medium containing recombinant chemokine served as a positive control. The supernatants of IFN-g-stimulated LC and splenic DC exhibited chemo- tactic activity to CXCR3 transfectant, which is mediated at least by CXCL10 and CXCL9. Mean (SD) (n ¼ 3). Data are representative of three independent experiments.

Article Snippet: Measurement of CXCL9, CXCL11, and CCL17 Culture supernatants were collected, stored at 201C, and subjected to the quantification of protein levels of CXCL9, CXCL11, and CCL17 by ELISA using commercially available mouse CXCL9, CXCL11, and CCL17 immunoassay kits (R&D Systems), respectively, according to the manufacturer’s instructions.

Techniques: Chemotaxis Assay, Transfection, Cell Culture, Activity Assay, Negative Control, Recombinant, Positive Control

Journal: Journal of Investigative Dermatology

Article Title: Differential Production of Th1- and Th2-Type Chemokines by Mouse Langerhans Cells and Splenic Dendritic Cells

doi: 10.1111/j.0022-202x.2004.23607.x

Figure Lengend Snippet: Figure 7 Regulation of T helper 1-type chemokines produced by La- ngerhans cells (LC) and splenic dendritic cells (DC). LC (&) and splenic DC (’) were purified and cultured (1.5 106 cells per mL per 200 mL in each well) for 48 h with or without various stimuli. The supe- rnatants were collected and the concentration of CXCL10 (a), CXCL9 (b), and CXCL11 (c) was measured by ELISA. CXCL10 production by LC was induced by interferon-g (IFN-g), interleukin (IL)-12, lipopoly- saccharide (LPS), Staphylococcus aureus Cowen 1 (SAC), and poly- inosinic–polycytidylic acid (Poly(I:C)). In the case of CXCL9 and CXCL11, only IFN-g induced their production by LC. In the case of splenic DC, the production of CXCL10, CXCL9, and CXCL11 was in- duced by IFN-g, IL-18, LPS, and Poly(I:C). Results are the mean (SD) (n ¼ 4). Significant increase (po0.05) compared with the unstimulated group. Data are representative of four independent experiments.

Article Snippet: Measurement of CXCL9, CXCL11, and CCL17 Culture supernatants were collected, stored at 201C, and subjected to the quantification of protein levels of CXCL9, CXCL11, and CCL17 by ELISA using commercially available mouse CXCL9, CXCL11, and CCL17 immunoassay kits (R&D Systems), respectively, according to the manufacturer’s instructions.

Techniques: Produced, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells

doi: 10.1101/2023.01.25.525561

Figure Lengend Snippet: (A) Culture supernatants collected from the upper and lower chambers of transwells with HUVEC monolayers were assayed for CCL2, CCL7 and CCL8 by ELISA. Data are from three experiments and bars indicate means +/- SEM. (B) Confocal microscopy images at ×40 magnification of non-permeabilized and permeabilized TNF-α + IFN-γ-stimulated HUVECs immunostained for CCL20, CCL5, CXCL9, CCL2, CCL7 or CCL8 (green) along with nuclear staining (blue) as described in Material and methods. The scale bars indicate 10 µm. Images are representative of three experiments. (C) CCR6 + CCR2 + cells were either left untreated or treated with CCL2 (100 ng/ml) just before and for 4 min after adding to the flow chambers, and CCL2-treated cells were used with or without treatment with the CCR2 antagonist, BMS22. Data points in the left panel show numbers of cells rolling, arrested and transmigrated, while the middle panel shows arrested cells as a percentage of cells rolling and the right panel shows transmigrated cells as a percentage of cells arresting. Each symbol shows data from cells from one donor, with four experiments using CCL2 and two experiments including the additional treatment with BMS22. Bars indicate means +/- SEM. p values were calculated using paired Student’s t tests, *, p<0.05; **, p<0.01; ***, p<0.001.

Article Snippet: Samples were incubated with primary antibodies against chemokines CCL2 (5 μg, MAB679) CCL5 (5 μg, MAB278), CCL7 (5 μg, MAB282), CCL8 (5 μg, MAB281), CCL20 (5μg, AF360), CXCL9 (5 μg, MAB392), mouse IgG 1 isotype control (5 μg, MAB002) and mouse IgG 2B isotype control (MAB004) all from R&D Systems, at RT for 2 h. Cells were washed three times with PBS before incubating with secondary antibody Alexa Fluor 488 (1:500, A-11017; Invitrogen) for 1 h and after washing three times with PBS, were counter-stained with DAPI (1μg/ml 62248; Invitrogen).

Techniques: Enzyme-linked Immunosorbent Assay, Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells

doi: 10.1101/2023.01.25.525561

Figure Lengend Snippet: (A) CCL2-CXCL9 chimera sequence with signal peptide sequence shown in green, CCL2-sequence in black and C-terminal GAG-binding sequence of CXCL9 in red (top). Confocal microscopy images of control or CCL2-CXCL9 chimera transduced, TNF- α-stimulated HUVECs immunostained for CCL2 (red) and DAPI (blue) as described in Materials and methods (bottom). (B) Numbers of CCR6 + CCR2 + cells, either untreated or treated with the CCR2 antagonist, BMS22, rolling, arrested and transmigrated on TNF-α-activated HUVECs transduced with either control virus or with virus encoding the CCL2-CXCL9 chimera. Middle panel shows arrested cells as a percentage of cells rolling and in the right panel transmigrated cells as a percentage of cells arresting. Each symbol shows data for cells from one donor, with cells from four donors in each treatment group except for the BMS22-treated cells with control virus-transduced HUVECs, where cells from three donors were used. Bars indicate means +/- SEM. p values were calculated using paired t tests, *, p<0.05; **, p<0.01; ***, p<0.001.

Article Snippet: Samples were incubated with primary antibodies against chemokines CCL2 (5 μg, MAB679) CCL5 (5 μg, MAB278), CCL7 (5 μg, MAB282), CCL8 (5 μg, MAB281), CCL20 (5μg, AF360), CXCL9 (5 μg, MAB392), mouse IgG 1 isotype control (5 μg, MAB002) and mouse IgG 2B isotype control (MAB004) all from R&D Systems, at RT for 2 h. Cells were washed three times with PBS before incubating with secondary antibody Alexa Fluor 488 (1:500, A-11017; Invitrogen) for 1 h and after washing three times with PBS, were counter-stained with DAPI (1μg/ml 62248; Invitrogen).

Techniques: Sequencing, Binding Assay, Confocal Microscopy, Control, Transduction, Virus

Journal: OncoImmunology

Article Title: Tumor-intrinsic RGS1 potentiates checkpoint blockade response via ATF3-IFNGR1 axis

doi: 10.1080/2162402x.2023.2279800

Figure Lengend Snippet: Figure 2. RGS1 is a modulator of IFNγ-JAK-STAT1 signaling. a Differentially expressed cancer-related gene sets (H_Hallmarks) with high RGS1 expression in the KIRC cohort, LUAD cohort, LUSC cohort, and SKCM cohort from TCGA. NES, normalized enrichment score. FDR q, false discovery rate q value. b – d GSEA output of genes in the HALLMARK_INTERFERON_GAMMA_RESPONSE by RGS1 high and low expression groups from the KIRC cohort (b), LUAD cohort (c) and LUSC cohort (d) in the TCGA database. ES, enrichment score. e – l Analysis of IFNγ-STAT1 signaling in 786O and Renca cells. Cell lysates of 786O and Renca cells stably expressing nc or shRGS1 (e, g) and Vector or RGS1-OE (i, k) were analyzed by western blotting using RGS1, STAT1, P-STAT1 (Y701), IFNGR1, and IRF1 antibodies. Tublin was used as an internal control. f, j IFNγ-inducible gene expression in 786O cells. mRNA expression of IRF1, IRF9, STAT1, and IFNGR1 were detected by real-time qPCR. Actin was used as an internal control. h, l IFNγ-inducible gene expression in Renca cells. mRNA expression of Irf1, Irf9, and Stat1 were detected by real-time qPCR. Actin was used as an internal control. Cells in e, f, i, j were stimulated with 10 ng/ml human recombinant IFNγ or 0.1% BSA negative control for 2 h. Cells in g, h, k, l were stimulated with 5 ng/ml mouse recombinant IFNγ or 0.1% BSA negative control for 2 h. m, n IFNγ-induced CXCL9 secretion. Renca (m) and LLC (n) cells were cultured in serum-free medium and treated with 5 ng/ml IFNγ for 24 h. The concentration of CXCL9 was analyzed using an ELISA kit. o, p Cell surface levels of IFNGR1 in nc or shRGS1 (o) and Vector or RGS1-OE (p) 786O cells (pre-gated with FSC-A vs. SSC-A, and FSC-A vs. FSC-H). Cells were treated with 10 ng/ml IFNγ for 2 h. Right, quantification of the mean fluorescence intensity (MFI). Unpaired t-test was performed with GraphPad Prism 9. All data are representative of three independent experiments. Data in the bar graphs represent mean ± S.D., n = 3. *p < .05, **p < .01, ***p < .001.

Article Snippet: The concentrations of mouse CXCL9/MIG, human cAMP level and the activity of PKA in the supernatants were analyzed using CXCL9 ELISA kit (R&D Systems, MCX900), cAMP ELISA Kit (Elabscience, E-EL-0056c) and PKA Colorimetric Activity Kit (Thermo Fisher scientific, EIAPKA) respectively, according to the manufacturer’s protocols.

Techniques: Expressing, Stable Transfection, Plasmid Preparation, Western Blot, Control, Gene Expression, Recombinant, Negative Control, Cell Culture, Concentration Assay, Enzyme-linked Immunosorbent Assay, Fluorescence

Journal: OncoImmunology

Article Title: Tumor-intrinsic RGS1 potentiates checkpoint blockade response via ATF3-IFNGR1 axis

doi: 10.1080/2162402x.2023.2279800

Figure Lengend Snippet: Figure 5. RGS1 is associated with T cell infiltration in RCC and NSCLC mouse models. a, d Representative images and quantification (right) of immunohistochemistry staining of IFNGR1, ATF3 and CXCL9 expression in harvested CTRL and ShRgs1 Renca (a) or LLC (d) subcutaneous tumor sections. The percentages of positively stained area (right) were analyzed using Image J software. Scale bar, 100 µm. b, e T cell infiltration and quantification (below) of Renca (b) or LLC (e) subcutaneous tumor. Paraffin-embedded tissue sections of murine tumors were immunohistochemically stained with antibodies against CD3, CD4, CD8 and PD1. The percentages of positively stained cells (below) were analyzed using Image J software. Scale bar, 100 µm. c, f Representative immunofluorescence images of CD8 and PD1 staining in ShRgs1 or CTRL Renca (c) or LLC (f) tumor sections. White arrows indicate merge of CD8 and PD1 fluorescence signals. Scale bar, 50 µm. Data in the graphs represent mean ± S.D, n = 5. *p < .05, **p < .01, ***p < .001.

Article Snippet: The concentrations of mouse CXCL9/MIG, human cAMP level and the activity of PKA in the supernatants were analyzed using CXCL9 ELISA kit (R&D Systems, MCX900), cAMP ELISA Kit (Elabscience, E-EL-0056c) and PKA Colorimetric Activity Kit (Thermo Fisher scientific, EIAPKA) respectively, according to the manufacturer’s protocols.

Techniques: Immunohistochemistry, Staining, Expressing, Software, Immunofluorescence, Fluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Cyclin G2 in macrophages triggers CTL-mediated antitumor immunity and antiangiogenesis via interferon-gamma

doi: 10.1186/s13046-022-02564-2

Figure Lengend Snippet: Cyclin G2 in macrophages regulates CTL chemotaxis and vascular endothelial cell tube formation via CXCL9. A CXCL9 levels in the supernatants of BMDMs from WT and Ccng2 −/− C57BL/6 mice treated with IFN-γ were determined by ELISA (representing 3 independent experiments). B , C CXCL9 levels in the supernatants of THP-1 stable cell lines (Nonsense, shcyclin G2#1, and shcyclin G2#2 and Vector and Flag-cyclin G2) treated with IFN-γ were determined by ELISA (representing 3 independent experiments). D , E CTL chemotaxis analyzed by treating conditioned medium from BMDMs isolated from WT and Ccng2 −/− C57BL/6 mice treated with or without recombinant CXCL9. Scale bar = 200 μm (representing 3 independent experiments). F Tube formation experiments showed the tube formation ability of SVEC4–10 cells treated with conditioned medium from BMDMs isolated from Ccng2 −/− C57BL/6 mice. The recombinant CXCL9 was added or not added to the conditioned medium. Scale bar = 500 μm (representing 3 independent experiments). G Tube formation experiments showed the tube formation ability of HUVECs treated with conditioned medium from a THP-1 stable cell line (shcyclin G2#1). The recombinant CXCL9 was added or not added to the conditioned medium. Scale bar = 200 μm (representing 3 independent experiments). (A–C, E–G) Data were analyzed with the unpaired Student’s t-test. Data are presented as the mean ± SD ** p < 0.01; *** p < 0.001; ns , not significant

Article Snippet: Cell culture supernatants were collected, and human and mouse CXCL9 levels in the supernatants were quantified using the human and mouse CXCL9 Enzyme-Linked Immunosorbent Assay (ELISA) Kits (E-EL-H6062 and E-EL-M0020c, Elabscience), respectively.

Techniques: Chemotaxis Assay, Enzyme-linked Immunosorbent Assay, Stable Transfection, Plasmid Preparation, Isolation, Recombinant