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recombinant mouse cxcl12 protein  (R&D Systems)


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    R&D Systems recombinant mouse cxcl12 protein
    Recombinant Mouse Cxcl12 Protein, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/sdf/Recombinant+Mouse+CXCL12%2FSDF-1+alpha+Protein/pm42021544-65-0-4
    Average 95 stars, based on 111 article reviews
    recombinant mouse cxcl12 protein - by Bioz Stars, 2026-11
    95/100 stars

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    Recombinant:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: BMP9 regulates the endothelial secretome to drive pulmonary hypertension
    Article Snippet: Monoclonal antibody to BMP9 (MAB3209), and biotinylated affinity purified antibodies to BMP9 (BAF3209) were obtained from R&D Systems. .. Recombinant CXCL12, PDGF-BB, TGFβ1, and anti-CXCL12 were purchased from R&D Systems. .. AMD3100 (CXCR4 inhibitor) was purchased from Selleck Chemicals.

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    Article Title: Diphlorethohydroxycarmalol inhibits Müller cell gliosis by disrupting CXCR4/CXCL12 interaction in violet-blue light-induced retinal phototoxicity.
    Article Snippet: .. In another in vitro model, CXCL12 induction was achieved by treating MIO-M1 cells with 20 nM recombinant CXCL12 protein (R&D Systems) for 1 d. Prior to the induction of CXCL12 in MIO-M1 cells, DPHC was pre-treated with the cells for 1 d. ..

    Article Title: Senolytic Treatment With Fisetin Reverses Age-Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12.
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine- 4M AM (DAR- 4M AM; Sigma- Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence- associated β- galactosidase (SA- β- Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460- SD- 050/CF) in sterile PBS was added back to plasma from young and old fisetin- treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle- treated mice (3210 pg/mL) for CXCL12 add- back experiments. .. 1 μg/ mL of LIT- 927 (Selleckchem, Cat. #S8813) (Xiong et al. 2022) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Article Title: Senolytic treatment with fisetin reverses age-related endothelial dysfunction partially mediated by SASP factor CXCL12
    Article Snippet: Endothelial cell nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine-4M AM (DAR-4M AM; Sigma-Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence-associated β-galactosidase (SA-β-Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460-SD-050/CF) in sterile PBS was added back to plasma from young and old fisetin-treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle-treated mice (3210 pg/ml) for CXCL12 add-back experiments. .. 1 μg/ml of LIT-927 (Selleckchem, Cat. #S8813) in DMSO was added to the old vehicle plasma for CXCL12 inhibtion experiments.

    Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines
    Article Snippet: The cells were then added to a TranswellTM plate insert with 5-μm pores (Corning Life Sciences, Corning, NY, USA #3421). .. Imaging medium (600 μL) with the same concentration of ATM-3507 or DMSO plus 100 nM recombinant human CXCL12 (R&D Systems, Minneapolis, MN, USA #460-SD) was added to the bottom chamber. ..

    Article Title: Senolytic Treatment With Fisetin Reverses Age‐Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine‐4M AM (DAR‐4M AM; Sigma‐Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence‐associated β‐galactosidase (SA‐β‐Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460‐SD‐050/CF) in sterile PBS was added back to plasma from young and old fisetin‐treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle‐treated mice (3210 pg/mL) for CXCL12 add‐back experiments. .. 1 μg/mL of LIT‐927 (Selleckchem, Cat. #S8813) (Xiong et al. ) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Purification:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    FACS:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Magnetic Cell Separation:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    Cell Isolation:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    Activity Assay:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. 153 2.5 Reagents and antibodies 154 The following reagents and materials were used in this study: Immunization 155 grade bovine type II collagen (Cat. No: 20021; Chondrex, Inc.), LUT (Cat. No: 156 B20888; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 157 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, 158 MA, USA), recombinant mouse C−X−C motif chemokine ligand 12 159 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 160 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free 161 (LEAF)TM purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San 162 Diego, CA, USA), ultra-LEAFTM purified anti-mouse CD28 antibody (Cat. No.: 163 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), 164 IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), 165 Jo urn al Pr e-p roo f 6 / 39 transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein 166 isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin 167 (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 168 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 169 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4+ T Cell Isolation 170 Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4+ T Cell Isolation Kit 171 (human) (Cat. No.: 130-096-533; Milteny Biotec), sodium L-lactate (Cat. No.: 71718; 172 Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: 173 BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit 174 (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), 175 mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits 176 (Cat. ..

    Article Title: Luteolin attenuates RA-associated chronic pain by targeting the LDHA/H3K9la/NFATC2 axis to suppress Th17 cell differentiation and central infiltration
    Article Snippet: .. The following reagents and materials were used in this study: immunization grade bovine type II collagen (Cat. No.: 20021; Chondrex, Inc.), LUT (Cat. No.: B20888 ; Shanghaiyuanye Bio-Technology Co., Ltd., Shanghai, China), anti-CD4 antibody (GK 1.5) (Cat. No.: 16-0041-86; Thermo Fisher Scientific Inc., Waltham, MA, USA), recombinant mouse C–X–C motif chemokine ligand 12 (CXCL12)/stromal cell-derived factor 1 alpha (SDF-1α) protein (Cat. No.: 460-SD-050; R&D Systems, Minneapolis, MN, USA), ultra-low endotoxin, azide-free (LEAF) purified anti-mouse CD3 antibody (Cat. No.: 100238; BioLegend, San Diego, CA, USA), ultra-LEAF purified anti-mouse CD28 antibody (Cat. No.: 102116; BioLegend), IL-1β (Cat. No.: 211-11B; PeproTech, Rocky Hill, NJ, USA), IL-23 (Cat. No.: 589002; BioLegend), IL-6 (Cat. No.: 216-16; PeproTech), transforming growth factor beta (TGF-β) (Cat. No.: 594509; BioLegend), fluorescein isothiocyanate (FITC)-CD3 (Cat. No.: 100203; BioLegend), allophycocyanin (APC)-CD4 (Cat. No.: 100515; BioLegend), phycoerythrin (PE)-IL-17A (Cat. No.: 506903; BioLegend), magnetic activated cell sorting (MACS) buffer (Cat. No.: 130-091-221; Miltenyi Biotec, Bergisch Gladbach, Germany), CD4 + T Cell Isolation Kit (mouse) (Cat. No.: 130-104-454; Miltenyi Biotec), CD4 + T Cell Isolation Kit (human) (Cat. No.: 130-096-533; Miltenyi Biotec), sodium l -lactate (Cat. No.: 71718; Sigma-Aldrich, St. Louis, MO, USA), Lactic Acid Content Assay Kit (Cat. No.: BC2230; Solarbio, Beijing, China), lactate dehydrogenase (LDH) Activity Assay Kit (Cat. No.: A020-2; Nanjing Jiancheng Bioengineering Institute, Nanjing, China), mouse IL-1β, IL-6, and TNF-α enzyme-linked immunosorbent assay (ELISA) kits (Cat. ..

    In Vitro:

    Article Title: Diphlorethohydroxycarmalol inhibits Müller cell gliosis by disrupting CXCR4/CXCL12 interaction in violet-blue light-induced retinal phototoxicity.
    Article Snippet: .. In another in vitro model, CXCL12 induction was achieved by treating MIO-M1 cells with 20 nM recombinant CXCL12 protein (R&D Systems) for 1 d. Prior to the induction of CXCL12 in MIO-M1 cells, DPHC was pre-treated with the cells for 1 d. ..

    Sterility:

    Article Title: Senolytic Treatment With Fisetin Reverses Age-Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12.
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine- 4M AM (DAR- 4M AM; Sigma- Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence- associated β- galactosidase (SA- β- Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460- SD- 050/CF) in sterile PBS was added back to plasma from young and old fisetin- treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle- treated mice (3210 pg/mL) for CXCL12 add- back experiments. .. 1 μg/ mL of LIT- 927 (Selleckchem, Cat. #S8813) (Xiong et al. 2022) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Article Title: Senolytic treatment with fisetin reverses age-related endothelial dysfunction partially mediated by SASP factor CXCL12
    Article Snippet: Endothelial cell nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine-4M AM (DAR-4M AM; Sigma-Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence-associated β-galactosidase (SA-β-Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460-SD-050/CF) in sterile PBS was added back to plasma from young and old fisetin-treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle-treated mice (3210 pg/ml) for CXCL12 add-back experiments. .. 1 μg/ml of LIT-927 (Selleckchem, Cat. #S8813) in DMSO was added to the old vehicle plasma for CXCL12 inhibtion experiments.

    Article Title: Senolytic Treatment With Fisetin Reverses Age‐Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine‐4M AM (DAR‐4M AM; Sigma‐Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence‐associated β‐galactosidase (SA‐β‐Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460‐SD‐050/CF) in sterile PBS was added back to plasma from young and old fisetin‐treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle‐treated mice (3210 pg/mL) for CXCL12 add‐back experiments. .. 1 μg/mL of LIT‐927 (Selleckchem, Cat. #S8813) (Xiong et al. ) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Clinical Proteomics:

    Article Title: Senolytic Treatment With Fisetin Reverses Age-Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12.
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine- 4M AM (DAR- 4M AM; Sigma- Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence- associated β- galactosidase (SA- β- Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460- SD- 050/CF) in sterile PBS was added back to plasma from young and old fisetin- treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle- treated mice (3210 pg/mL) for CXCL12 add- back experiments. .. 1 μg/ mL of LIT- 927 (Selleckchem, Cat. #S8813) (Xiong et al. 2022) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Article Title: Senolytic treatment with fisetin reverses age-related endothelial dysfunction partially mediated by SASP factor CXCL12
    Article Snippet: Endothelial cell nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine-4M AM (DAR-4M AM; Sigma-Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence-associated β-galactosidase (SA-β-Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460-SD-050/CF) in sterile PBS was added back to plasma from young and old fisetin-treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle-treated mice (3210 pg/ml) for CXCL12 add-back experiments. .. 1 μg/ml of LIT-927 (Selleckchem, Cat. #S8813) in DMSO was added to the old vehicle plasma for CXCL12 inhibtion experiments.

    Article Title: Senolytic Treatment With Fisetin Reverses Age‐Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine‐4M AM (DAR‐4M AM; Sigma‐Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence‐associated β‐galactosidase (SA‐β‐Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460‐SD‐050/CF) in sterile PBS was added back to plasma from young and old fisetin‐treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle‐treated mice (3210 pg/mL) for CXCL12 add‐back experiments. .. 1 μg/mL of LIT‐927 (Selleckchem, Cat. #S8813) (Xiong et al. ) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Concentration Assay:

    Article Title: Senolytic Treatment With Fisetin Reverses Age-Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12.
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine- 4M AM (DAR- 4M AM; Sigma- Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence- associated β- galactosidase (SA- β- Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460- SD- 050/CF) in sterile PBS was added back to plasma from young and old fisetin- treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle- treated mice (3210 pg/mL) for CXCL12 add- back experiments. .. 1 μg/ mL of LIT- 927 (Selleckchem, Cat. #S8813) (Xiong et al. 2022) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Article Title: Senolytic treatment with fisetin reverses age-related endothelial dysfunction partially mediated by SASP factor CXCL12
    Article Snippet: Endothelial cell nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine-4M AM (DAR-4M AM; Sigma-Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence-associated β-galactosidase (SA-β-Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460-SD-050/CF) in sterile PBS was added back to plasma from young and old fisetin-treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle-treated mice (3210 pg/ml) for CXCL12 add-back experiments. .. 1 μg/ml of LIT-927 (Selleckchem, Cat. #S8813) in DMSO was added to the old vehicle plasma for CXCL12 inhibtion experiments.

    Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines
    Article Snippet: The cells were then added to a TranswellTM plate insert with 5-μm pores (Corning Life Sciences, Corning, NY, USA #3421). .. Imaging medium (600 μL) with the same concentration of ATM-3507 or DMSO plus 100 nM recombinant human CXCL12 (R&D Systems, Minneapolis, MN, USA #460-SD) was added to the bottom chamber. ..

    Article Title: Senolytic Treatment With Fisetin Reverses Age‐Related Endothelial Dysfunction Partially Mediated by SASP Factor CXCL12
    Article Snippet: EC nitric oxide (NO) and mitochondrial superoxide bioactivity were assessed after 2 h plasma incubation by staining with the fluorescent probe Hoechst (nuclei stain; Thermo Fisher) and either 10 μM diaminorhodamine‐4M AM (DAR‐4M AM; Sigma‐Aldrich; to quantify NO production) for 45 min or 5 μM MitoSOX (Thermo Fisher; to quantify mitochondrial superoxide bioactivity) for 30 min. Senescence‐associated β‐galactosidase (SA‐β‐Gal) staining and RNA extractions for gene expression were performed after 24 h of plasma exposure. .. Recombinant mouse CXCL12 protein (R&D Systems, Cat. #460‐SD‐050/CF) in sterile PBS was added back to plasma from young and old fisetin‐treated mice, such that the concentration of CXCL12 matched the average levels in old vehicle‐treated mice (3210 pg/mL) for CXCL12 add‐back experiments. .. 1 μg/mL of LIT‐927 (Selleckchem, Cat. #S8813) (Xiong et al. ) in DMSO was added to the old vehicle plasma for CXCL12 inhibition experiments.

    Imaging:

    Article Title: The tropomyosin 3.1/3.2 inhibitor ATM-3507 alters B-cell actin dynamics and impairs the growth and motility of diffuse large B-cell lymphoma cell lines
    Article Snippet: The cells were then added to a TranswellTM plate insert with 5-μm pores (Corning Life Sciences, Corning, NY, USA #3421). .. Imaging medium (600 μL) with the same concentration of ATM-3507 or DMSO plus 100 nM recombinant human CXCL12 (R&D Systems, Minneapolis, MN, USA #460-SD) was added to the bottom chamber. ..



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    ( A ) Cell-cell communication among clusters determined by CellChat. ( B ) GO term enrichment analysis in Fibroblast2. ( C ) Chemokine expression in each cluster. ( D ) CXCL1 and <t>CXCL12</t> expression by IL-6/soluble IL-6Rα stimulation at protein levels in FLS in AIA. N = 5 from five independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( E ) CXCR2 expression in human RA synovial samples (phs001457.v1.p1). ( F ) CXCL1 expression by IL-6/soluble IL-6Rα stimulation at protein levels in human RA FLS. N = 9 from four independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( G ) The expression of chemokine receptor Cxcr2 in AIA. ( H ) The expression of CXCR2 in peripheral blood (PB) and synovial fluid (SF) neutrophils in RA ( GSE154474 and GSE116899 ). ( I to L ) The chemokine receptor antagonist experiment in the stress-induced AIA model. N = 9 to 10 from three independent experiments. (I) Ankle thickness after the intervention. (J) Ankle thickness at day 24. (K) Representative images of ankles. (L) Cumulative data of the histological score. P values were determined by unpaired t test.
    Human Cxcl12 Sdf 1 Elisa Kit, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; <t>CXCR4-CXCL12</t> interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .
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    A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; <t>CXCR4-CXCL12</t> interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .
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    A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; <t>CXCR4-CXCL12</t> interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .
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    Image Search Results


    ( A ) Cell-cell communication among clusters determined by CellChat. ( B ) GO term enrichment analysis in Fibroblast2. ( C ) Chemokine expression in each cluster. ( D ) CXCL1 and CXCL12 expression by IL-6/soluble IL-6Rα stimulation at protein levels in FLS in AIA. N = 5 from five independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( E ) CXCR2 expression in human RA synovial samples (phs001457.v1.p1). ( F ) CXCL1 expression by IL-6/soluble IL-6Rα stimulation at protein levels in human RA FLS. N = 9 from four independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( G ) The expression of chemokine receptor Cxcr2 in AIA. ( H ) The expression of CXCR2 in peripheral blood (PB) and synovial fluid (SF) neutrophils in RA ( GSE154474 and GSE116899 ). ( I to L ) The chemokine receptor antagonist experiment in the stress-induced AIA model. N = 9 to 10 from three independent experiments. (I) Ankle thickness after the intervention. (J) Ankle thickness at day 24. (K) Representative images of ankles. (L) Cumulative data of the histological score. P values were determined by unpaired t test.

    Journal: Science Advances

    Article Title: Fibroblast-like synoviocytes exacerbate autoimmune arthritis upon psychological stress in an IL-6–dependent manner

    doi: 10.1126/sciadv.aec9257

    Figure Lengend Snippet: ( A ) Cell-cell communication among clusters determined by CellChat. ( B ) GO term enrichment analysis in Fibroblast2. ( C ) Chemokine expression in each cluster. ( D ) CXCL1 and CXCL12 expression by IL-6/soluble IL-6Rα stimulation at protein levels in FLS in AIA. N = 5 from five independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( E ) CXCR2 expression in human RA synovial samples (phs001457.v1.p1). ( F ) CXCL1 expression by IL-6/soluble IL-6Rα stimulation at protein levels in human RA FLS. N = 9 from four independent experiments. P values were determined by Friedman test followed by Dunn’s test. ( G ) The expression of chemokine receptor Cxcr2 in AIA. ( H ) The expression of CXCR2 in peripheral blood (PB) and synovial fluid (SF) neutrophils in RA ( GSE154474 and GSE116899 ). ( I to L ) The chemokine receptor antagonist experiment in the stress-induced AIA model. N = 9 to 10 from three independent experiments. (I) Ankle thickness after the intervention. (J) Ankle thickness at day 24. (K) Representative images of ankles. (L) Cumulative data of the histological score. P values were determined by unpaired t test.

    Article Snippet: Mouse CXCL1 and CXCL12 levels of the culture supernatant were determined using a Mouse CXCL1 ELISA kit (Proteintech, Rosemont, IL) or a Mouse CXCL12 ELISA Kit (Proteintech).

    Techniques: Analysis, Expressing

    A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; CXCR4-CXCL12 interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .

    Journal: Nature Communications

    Article Title: Neoadjuvant chemotherapy interferes with tertiary lymphoid structure formation in Epstein-Barr virus-associated gastric cancer

    doi: 10.1038/s41467-026-77546-5

    Figure Lengend Snippet: A UMAP embedding revealing six distinct subtypes of neutrophils identified in scRNA-seq samples ( n = 21). B KEGG pathway enrichments (fold enrichment) in different neutrophil subtypes ( n = 21). C Pseudotime cell trajectory of neutrophils ( n = 21) colored by subtypes (left) and dynamic expression of key chemokine receptors along the trajectory (right). D Dot plot showing expression levels and percentages of key chemokine receptors across the six neutrophil subtypes ( n = 21). E UMAP plot (left) showing re-clustering of neutrophils ( n = 21) into two functional states: aging neutrophils (Neu_aging) and inflammatory neutrophils (Neu_inflam). Feature plots (right) showing expression of aging markers SELL (CD62L, downregulated) and CXCR4 (upregulated). F Spatial feature maps ( n = 11), from top to bottom: spatial regions and regional TLSs; CXCR4-CXCL12 interaction scores; aging neutrophil abundance; and CXCR4 expression derived from aging neutrophils. Red arrows indicate co-localization of aging neutrophils at the periphery of IM-TLSs. G Bubble plot showing CXCL signaling communication probabilities between neutrophil subtypes and other microenvironmental populations in scRNAseq samples ( n = 21). Significance assessed by one-sided permutation test (100 permutations). H Representative mIHC images showing co-localization of aging neutrophils (CD66b + CXCR4+) and CXCL12-expressing stromal cells (α-SMA + CXCL12+) around a TLS (CD20 + B cells). Images are representative of 216 independent patient samples examined. Scale bar, 100 μm. I. Schematic of the in vitro Transwell assay evaluating CXCL12-mediated chemotaxis of aging neutrophils (Created in BioRender. Bi, J. (2026) https://BioRender.com/bhu0jn4 ). Representative FACS histograms of CXCR4 expression on neutrophils indirectly co-cultured with J EBV-positive SNU719 or K EBV-negative AGS cells, with or without 5-FU (NC vs. 5-FU). FACS gating strategy shown in Supplementary Fig. . Transwell migration assays ( n = 3) showing the chemotactic effect of CXCL12 on neutrophils co-cultured with L SNU719 or M AGS across treatment groups. FACS gating strategy shown in Supplementary Fig. . Two-sided Student’s t test with Bonferroni correction for multiple comparisons (ns, P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001). Exact P values are shown in .

    Article Snippet: The markers incorporated in Panel 3 were PanCK (1:2000, Clone PDH09-10, Cat# HA601138 , HUABIO), BZLF1 (1:400, Clone BZ1, Cat#sc-53904, SANTA CRUZ), CD20 (1:2000, Clone #06, Cat#11007-MM06, Sino Biological), CD23 (1:500, Clone PD00-03, Cat#HA721139, HUABIO), CD66b (1:20,000, Clone EPR25354-2, Cat#ab300122, Abcam), MPO(1:200, Polyclonal, Cat#AF3667, R&D), CitH3(1:200, Polyclonal, Cat#ab5103, Abcam), CXCR4 (1:500, Clone UMB2, Cat#ab124824, Abcam), CXCL12 (1:100, Polyclonal, Cat#17402-1-AP, proteintech), αSMA (1:8000, Polyclonal, Cat#14395-1-AP, proteintech), and DAPI.

    Techniques: Expressing, Functional Assay, Derivative Assay, In Vitro, Transwell Assay, Chemotaxis Assay, Cell Culture, Migration

    Under the stressful conditions induced by 5-FU-based neoadjuvant chemotherapy, latent EBV undergoes lytic reactivation in cancer cells. Upon reactivation and cancer cell lysis, EBV releases viral pathogen-associated molecular patterns (PAMPs), including viral DNA, RNA, and proteins, which are subsequently recognized by pattern recognition receptors (PRRs), such as TLR7, on tumor-infiltrating neutrophils. This recognition triggers neutrophil phenotypic transformation toward an aging phenotype characterized by upregulated CXCR4 expression and enhanced NET-formation capacity. These aging neutrophils are recruited to the periphery of TLS via CXCR4-CXCL12 chemotaxis, where they release NETs that form a physical barrier isolating TLS as immunological “islands” This NET-mediated barrier prevents the continuous recruitment of antigen-presenting cells and disrupts TLS maturation and maintenance, ultimately contributing to poor prognosis in EBVaGC patients. (Created in BioRender. Bi, J. (2026) https://BioRender.com/3a9ekcx ).

    Journal: Nature Communications

    Article Title: Neoadjuvant chemotherapy interferes with tertiary lymphoid structure formation in Epstein-Barr virus-associated gastric cancer

    doi: 10.1038/s41467-026-77546-5

    Figure Lengend Snippet: Under the stressful conditions induced by 5-FU-based neoadjuvant chemotherapy, latent EBV undergoes lytic reactivation in cancer cells. Upon reactivation and cancer cell lysis, EBV releases viral pathogen-associated molecular patterns (PAMPs), including viral DNA, RNA, and proteins, which are subsequently recognized by pattern recognition receptors (PRRs), such as TLR7, on tumor-infiltrating neutrophils. This recognition triggers neutrophil phenotypic transformation toward an aging phenotype characterized by upregulated CXCR4 expression and enhanced NET-formation capacity. These aging neutrophils are recruited to the periphery of TLS via CXCR4-CXCL12 chemotaxis, where they release NETs that form a physical barrier isolating TLS as immunological “islands” This NET-mediated barrier prevents the continuous recruitment of antigen-presenting cells and disrupts TLS maturation and maintenance, ultimately contributing to poor prognosis in EBVaGC patients. (Created in BioRender. Bi, J. (2026) https://BioRender.com/3a9ekcx ).

    Article Snippet: The markers incorporated in Panel 3 were PanCK (1:2000, Clone PDH09-10, Cat# HA601138 , HUABIO), BZLF1 (1:400, Clone BZ1, Cat#sc-53904, SANTA CRUZ), CD20 (1:2000, Clone #06, Cat#11007-MM06, Sino Biological), CD23 (1:500, Clone PD00-03, Cat#HA721139, HUABIO), CD66b (1:20,000, Clone EPR25354-2, Cat#ab300122, Abcam), MPO(1:200, Polyclonal, Cat#AF3667, R&D), CitH3(1:200, Polyclonal, Cat#ab5103, Abcam), CXCR4 (1:500, Clone UMB2, Cat#ab124824, Abcam), CXCL12 (1:100, Polyclonal, Cat#17402-1-AP, proteintech), αSMA (1:8000, Polyclonal, Cat#14395-1-AP, proteintech), and DAPI.

    Techniques: Lysis, Transformation Assay, Expressing, Chemotaxis Assay