antibodies against ip 10  (Bio-Rad)

Journal: bioRxiv

Article Title: Microengineered transplantation of human solid tumors for in vitro studies of CAR T immunotherapy

doi: 10.1101/2025.04.25.650709

Figure Lengend Snippet: a, Conceptual diagram illustrating pairwise mapping of directional ligand-receptor interactions between different cell types in the engineered tumor model. b,c, Chord diagrams of cell pair-specific ligand-receptor interactions with adjusted p-values < 0.05 and total mean expression > 0.35. d, Visualization of interactions between CAR T cells (CAR Ts) and endothelial cells (Endos) mediated by CXCR3/DPP4-CXCL10/CXCL11. e, Violin plots comparing the expression of these mediators across all cell types. f, Experimental timeline for CAR T cell infusion and drug treatment. g, Representative confocal micrographs of single tumors treated with CAR T cells and different concentrations of LAF237. Blood vessels are not shown in these images. Scale bars, 250 μm. h, Representative images of CAR T cell-infused tumors at Day 26. Scale bars, 200 µm. i, Quantification and comparison of tumor area (top) and CAR T cell-occupied hydrogel area (bottom). Shaded in grey is a period of CAR T cell infusion and daily LAF237 treatment. Data are presented as mean ± SEM (n ≥ 3). j,k, Quantification of CXCL10 and CXCL11 levels ( j ) and the activity of DPP4 ( k ) in device effluent. Shaded in grey is a period of CAR T cell infusion and daily LAF237 treatment. Data are presented as mean ± SEM in ( j ) and as mean ± SD in ( k ) (n ≥ 3). CAR T cells derived from three healthy donors were tested in this study.

Article Snippet: The membranes were blocked with 5% skim milk for 1 hour at room temperature, followed by overnight incubation at 4°C with primary antibodies against CXCL10 (MAB266-100, R&D systems).

Techniques: Expressing, Comparison, Activity Assay, Derivative Assay

Journal: Cell death & disease

Article Title: MLKL signaling regulates macrophage polarization in acute pancreatitis through CXCL10.

doi: 10.1038/s41419-023-05655-w

Figure Lengend Snippet: Fig. 6 RNA sequencing was performed to identify the differentially expressed genes between WT and Mlkl-/- mouse-derived, cerulein- treated pancreatic acinar cells. CXCL10 was the most significantly upregulated gene in pancreatic acinar cells from WT mice and was validated in cell and animal experiments. A Schematic diagram for preparing samples for RNA sequencing. B Number of differentially expressed genes (DEGs) at different fold changes. C Heatmap of the top 20 upregulated and downregulated genes. D Volcano plot of DEGs. E PCA plot. F mRNA levels of Cxcl10 in pancreatic acinar cells isolated from WT and Mlkl-/- mice. Data were normalized to GAPDH (n = 6). G Levels of CXCL10 in the supernatant of pancreatic acinar cells isolated from WT and Mlkl-/- mice (n = 6). H Serum CXCL10 levels in WT and Mlkl-/- mice (n = 7). I Immunofluorescence staining of amylase (green) colocalized with CXCL10 (red) in the pancreas of WT and Mlkl-/- mice (n = 3). Values are shown as the means ± SEMs. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. WT wild type, NS normal saline, AP acute pancreatitis, CR cerulein, MFI mean fluorescence intensity.

Article Snippet: The neutralizing antibody against CXCL10 (GTX31179) and rabbit IgG isotype control (GTX35035) for in vitro experiments were purchased from GeneTex (Shanghai, China), while the neutralizing antibody against CXCL10 (MAB466) and rat IgG isotype control (6-001-F) for in vivo experiments were purchased from R&D Systems (Shanghai, China).

Techniques: RNA Sequencing, Derivative Assay, Isolation, Staining, Saline

Journal: Cell death & disease

Article Title: MLKL signaling regulates macrophage polarization in acute pancreatitis through CXCL10.

doi: 10.1038/s41419-023-05655-w

Figure Lengend Snippet: Fig. 7 Rabbit anti-mouse CXCL10 neutralizing antibody (2 μg/mL) was added to the conditioned medium from WT mouse-derived, cerulein-treated pancreatic acinar cells. Rabbit IgG was added as an isotype control, and neutralization of CXCL10 impaired its ability to promote macrophage polarization toward M1. A Schematic diagram of constructing an indirect coculture system and preparing conditioned medium with CXCL10 neutralization. B Immunofluorescence staining of iNOS (red) and CD206 (green) in peritoneal macrophages (n = 3). C, D mRNA levels of M1 macrophage markers (Inos, Cd86, Il-12a, Tnf-α) and M2 macrophage markers (Cd163, Cd206, Ym-1) in peritoneal macrophages. Data were normalized to GAPDH (n = 6–8). Values are shown as the means ± SEMs. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. WT wild type, Con control, CR cerulein, CM conditioned medium.

Article Snippet: The neutralizing antibody against CXCL10 (GTX31179) and rabbit IgG isotype control (GTX35035) for in vitro experiments were purchased from GeneTex (Shanghai, China), while the neutralizing antibody against CXCL10 (MAB466) and rat IgG isotype control (6-001-F) for in vivo experiments were purchased from R&D Systems (Shanghai, China).

Techniques: Derivative Assay, Control, Neutralization, Staining

Journal: Cell death & disease

Article Title: MLKL signaling regulates macrophage polarization in acute pancreatitis through CXCL10.

doi: 10.1038/s41419-023-05655-w

Figure Lengend Snippet: Fig. 8 Rat anti-mouse CXCL10 dissolved in saline was administered after the first cerulein injection in C57BL/6J mice (20 μg/each), while rat IgG was injected as an isotype control. In vivo neutralization of CXCL10 alleviated AP in mice by reducing macrophage M1 polarization. A HE staining and pancreatic pathological scores of the pancreas (n = 6). B Serum amylase (n = 6). C, D Serum TNF-α and IL-6 (n = 6). E Immunofluorescence staining of F4/80 (green) colocalized with iNOS (red) or CD206 (red) in the pancreas of mice and the percentage of macrophages (n = 6). Values are shown as the means ± SEMs. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. NS normal saline, AP acute pancreatitis.

Article Snippet: The neutralizing antibody against CXCL10 (GTX31179) and rabbit IgG isotype control (GTX35035) for in vitro experiments were purchased from GeneTex (Shanghai, China), while the neutralizing antibody against CXCL10 (MAB466) and rat IgG isotype control (6-001-F) for in vivo experiments were purchased from R&D Systems (Shanghai, China).

Techniques: Saline, Injection, Control, In Vivo, Neutralization, Staining

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins induce CXCL10 in a cell type-specific manner. ( A ) SDS-polyacrylamide gel electrophoresis of recombinant human galectins (rhGal) stained with Coomassie Blue. Molecular weights of protein standards are indicated on the left. The hemagglutination assay was performed using serial two-fold dilutions of the recombinant protein. The bottom row contains protein incubated with β-lactose, a competitive carbohydrate inhibitor of galectin binding. ( B ) Cultures of human corneal fibroblasts, human corneal epithelial cells, THP-1 monocytes and human umbilical vein endothelial cells (HUVECs) were incubated with 50 µg/mL rhGal-1, -3 and -8 or 50 µg/mL IFN-γ. Quantitative real-time PCR was used to determine the levels of CXCL10 mRNA after 6 h of incubation ( n = 3 independent experiments), whereas ELISA was used to determine the levels of CXCL10 protein in cell culture supernatants after 24 h of incubation ( n = 6 independent experiments). The data represent the mean ± SEM.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Polyacrylamide Gel Electrophoresis, Recombinant, Staining, Hemagglutination Assay, Incubation, Binding Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Cells

Article Title: Induction of CXCL10-Mediated Cell Migration by Different Types of Galectins

doi: 10.3390/cells10020274

Figure Lengend Snippet: Galectins differentially dictate fibroblast-dependent immune cell chemotaxis. ( A ) Recombinant human galectin (rhGal)-1, -3 and -8 were incubated with fibroblasts for 24 h. The conditioned media were pre-cleared with β-lactose Sepharose beads three times to remove galectins prior to chemotaxis assays. The presence of galectins in pre-cleared conditioned media was evaluated by immunoblotting. ( B ) Quantification by flow cytometry of the migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with increasing concentrations of recombinant galectins or BSA ( n = 3 independent experiments performed in duplicate). ( C ) Migration of CFSE-labeled THP-1 and Jurkat cells to conditioned media from fibroblasts treated for 24 h with 50 μg/mL of recombinant galectins or BSA. A CXCL10 neutralizing antibody was added to the conditioned media in neutralization studies ( n = 4–5 independent experiments in duplicate). The data in ( B ) represent the mean ± SEM. The box and whisker plots show the 25 and 75 percentiles (box), the median, and the minimum and maximum data values (whiskers). Significance was determined using Student’s t test. ** p < 0.01.

Article Snippet: A mouse monoclonal antibody against CXCL10 (1.2 μg/mL; MAB266, R&D Systems) was added to the lower compartment of the chamber in neutralization studies.

Techniques: Chemotaxis Assay, Recombinant, Incubation, Western Blot, Flow Cytometry, Migration, Labeling, Neutralization, Whisker Assay