Journal:

Article Title: Pleiotropic effects of CXC chemokines in gastric carcinoma: differences in CXCL8 and CXCL1 expression between diffuse and intestinal types of gastric carcinoma

doi: 10.1111/j.1365-2249.2003.02305.x

Figure Lengend Snippet: CXCL8 and CXCL1 expression by gastric carcinoma cells, endothelial cells and neutrophils by immunohistochemistry. (a–c) CXCL8 was expressed by carcinoma cells (arrowheads) in all patients with gastric carcinoma. Both the number of CXCL8-expressing tumour cells as well as the intensity of CXCL8 expression by tumour cells was significantly stronger in diffuse- (a) compared to intestinal (b) type-gastric carcinoma. The inset in (a) shows expression of CXCL8 by endothelial cells in the tumour tissue. The isotype control for CXCL8 is completely negative (c). (d–f) CXCL1 was expressed by carcinoma cells (arrowheads) of diffuse-type gastric carcinoma (d), whereas expression of CXCL1 was not detected in most cases of intestinal-type gastric carcinoma cells (arrowheads). CXCL1 expressing tumour-infiltrating neutrophils (arrows) serve as an internal positive control (e). Inset in (e) shows negative carcinoma cells (arrowheads) and positive neutrophils (arrows) in more detail.The isotype control for CXCL1 is completely negative (f). Magnification (a–d,f) ×500; (e) ×1000.

Article Snippet: For immunohistological analyses the following monoclonal and polyclonal antibodies were used at the dilutions indicated: mouse monoclonal anti-CD3 (1 : 500, Dako, Hamburg, Germany), mouse monoclonal anti-CD68 (1 : 8000, clone KIM6, gift from the Institute of Pathology, Kiel, Germany), mouse monoclonal anti-CD34 (1 : 20, Immunotech, Marseille, France), mouse monoclonal anti-CXCL8 (1 : 50, Bender Medical Systems, Vienna, Austria, clone NAPII); mouse monoclonal anti-CXCL1 (1 : 25, R&D Systems, Wiesbaden, Germany, clone 20326·1); rabbit polyclonal anti-CXCL10 (1 : 500, Pepro Tech EC Ltd, London, UK); rabbit polyclonal anti-CXCL9 (1 : 800, Pepro Tech); mouse monoclonal anti-IFN- γ (1 : 50, R&D Systems, clone 42705·111); mouse monoclonal anti-CXCR1 (1 : 100, R&D Systems, clone 42705·111); mouse monoclonal anti-CXCR2 (1 : 100, R&D Systems, clone 48311·211); mouse monoclonal anti-CXCR3 (1 : 4000, R&D Systems, clone 49801·111).

Techniques: Expressing, Immunohistochemistry, Positive Control

Journal:

Article Title: Pleiotropic effects of CXC chemokines in gastric carcinoma: differences in CXCL8 and CXCL1 expression between diffuse and intestinal types of gastric carcinoma

doi: 10.1111/j.1365-2249.2003.02305.x

Figure Lengend Snippet: Expression of the chemokine receptors CXCR1, CXCR2 and CXCR3 in gastric carcinoma determined by immunohistochemistry. (a) CXCR1 was expressed strongly on neutrophils and some mononuclear cells, representing lymphocytes and macrophages infiltrating the tumour stroma. This was shown examplarily in an intestinal-type gastric carcinoma. (b–f): Carcinoma cells expressed the CXCR1 receptor (b,c): intestinal-type gastric carcinoma; (d): diffuse-type gastric carcinoma; carcinoma cells marked with arrowheads; (e,f): negative isotype controls for CXCR1). Expression was pronounced at the invasive edge (arrowheads) of the carcinoma (b). (g) Carcinoma cells expressed the CXCR2 receptor as examplarily shown in intestinal-type gastric carcinoma. Our data suggest that the corresponding chemokines CXCL8 and CXCL1 may act on carcinoma cells themselves by an autocrine or paracrine mechanism. (h) CXCR3, the receptor for CXCL10 and CXCL9, was expressed on tumour-infiltrating lymphocytes. Arrowheads mark gastric carcinoma glands. Magnification (a,c–f) ×1000; (b) ×250; (g,h) ×500.

Article Snippet: For immunohistological analyses the following monoclonal and polyclonal antibodies were used at the dilutions indicated: mouse monoclonal anti-CD3 (1 : 500, Dako, Hamburg, Germany), mouse monoclonal anti-CD68 (1 : 8000, clone KIM6, gift from the Institute of Pathology, Kiel, Germany), mouse monoclonal anti-CD34 (1 : 20, Immunotech, Marseille, France), mouse monoclonal anti-CXCL8 (1 : 50, Bender Medical Systems, Vienna, Austria, clone NAPII); mouse monoclonal anti-CXCL1 (1 : 25, R&D Systems, Wiesbaden, Germany, clone 20326·1); rabbit polyclonal anti-CXCL10 (1 : 500, Pepro Tech EC Ltd, London, UK); rabbit polyclonal anti-CXCL9 (1 : 800, Pepro Tech); mouse monoclonal anti-IFN- γ (1 : 50, R&D Systems, clone 42705·111); mouse monoclonal anti-CXCR1 (1 : 100, R&D Systems, clone 42705·111); mouse monoclonal anti-CXCR2 (1 : 100, R&D Systems, clone 48311·211); mouse monoclonal anti-CXCR3 (1 : 4000, R&D Systems, clone 49801·111).

Techniques: Expressing, Immunohistochemistry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deletion of C-C Motif Chemokine Ligand 5 Worsens Invariant Natural Killer T-Cell–Mediated Hepatitis via Compensatory Up-regulation of CXCR2–Related Chemokine Activity

doi: 10.1016/j.jcmgh.2018.12.009

Figure Lengend Snippet: Ccl5 deficiency synergistically up-regulates neutrophil infiltration in peripheral blood. ( A ) Representative flow cytometry profiles showing percentages of neutrophils (CD11b high Gr1 high ) gated in CD45 + peripheral blood cells from each group. ( B ) Proportion of neutrophils was shown, and total number of neutrophils was calculated. ( C ) Mice were treated with α-Galcer for 3 hours, liver tissues were collected, and total RNA was isolated and subjected to real-time PCR analysis of Cxcl1, Cxcl2, Cxcr2, Icam-1, E-selectin, and P-selectin . ( D ) Sera were collected to measure CXCL1 levels with ELISA kit. ( E ) Sera were collected to measure CXCL2 levels with ELISA kit. * P < .05.

Article Snippet: Antibodies used in immunohistochemistry staining included MPO (cat number: 901-023-100510; Biocare Medical, Concord, CA) and CXCL1 (cat number: BS-10234R; Bioss, Beijing, China).

Techniques: Flow Cytometry, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deletion of C-C Motif Chemokine Ligand 5 Worsens Invariant Natural Killer T-Cell–Mediated Hepatitis via Compensatory Up-regulation of CXCR2–Related Chemokine Activity

doi: 10.1016/j.jcmgh.2018.12.009

Figure Lengend Snippet: Up-regulation of Cxcl1 mRNA expression in liver and hepatocytes from α-Galcer–treated Ccl5 -/- mice. Mice were treated with α-Galcer for 3 hours, and various organs were collected and subjected to quantitative real-time PCR analysis of ( A ) Cxcl1 and ( B ) Cxcl2 mRNA. Hepatocytes and NPCs were isolated and subjected to real-time PCR analysis to determine levels of ( C ) Cxcl1 and ( D ) Cxcl2 mRNA. ( E ) Representative CXCL1 staining of liver tissue sections. ( F ) Quantitative real-time PCR analysis of Ccr1 , Ccr3, and Ccr5 in liver tissue from 3-hour α-Galcer treatment. ( G ) Quantitative real-time PCR analysis of Ccr1 , Ccr3, and Ccr5 in hepatocytes isolated from α-Galcer–treated mice for 3 hours. ( H ) Quantitative real-time PCR analysis of peroxisome proliferative-activated receptor-γ in liver tissue from 3-hour α-Galcer treatment. * P < .05, ** P < .01.

Article Snippet: Antibodies used in immunohistochemistry staining included MPO (cat number: 901-023-100510; Biocare Medical, Concord, CA) and CXCL1 (cat number: BS-10234R; Bioss, Beijing, China).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deletion of C-C Motif Chemokine Ligand 5 Worsens Invariant Natural Killer T-Cell–Mediated Hepatitis via Compensatory Up-regulation of CXCR2–Related Chemokine Activity

doi: 10.1016/j.jcmgh.2018.12.009

Figure Lengend Snippet: Augmented neutrophil recruitment is dependent on CXCL1 and CXCR2 mediated migration in an in vitro system. ( A ) Purity of MACS based isolation of neutrophil was verified by flow cytometry analysis. ( B ) Neutrophils isolated from α-Galcer–treated liver tissue were analyzed for their migration in response to 0-hour serum, 3-hour serum, or with anti-CXCL1 ( D ) or SB225002 ( F ). ( C , E , and G ) In magnification of ×100, 8–10 images of each group were taken randomly, and adjusted count was analyzed by Image-Pro Plus. * P < .05, ** P < .01, *** P < .001.

Article Snippet: Antibodies used in immunohistochemistry staining included MPO (cat number: 901-023-100510; Biocare Medical, Concord, CA) and CXCL1 (cat number: BS-10234R; Bioss, Beijing, China).

Techniques: Migration, In Vitro, Isolation, Flow Cytometry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Deletion of C-C Motif Chemokine Ligand 5 Worsens Invariant Natural Killer T-Cell–Mediated Hepatitis via Compensatory Up-regulation of CXCR2–Related Chemokine Activity

doi: 10.1016/j.jcmgh.2018.12.009

Figure Lengend Snippet: Blockade of CXCL1-CXCR2 axis diminished the enhanced hepatitis in Ccl5 -/- mice. ( A ) ALT and ( B ) AST were measured in serum from anti-CXCL1 and isotype control pretreatment mice, subsequently administered α-Galcer for 24 hours. WT and Ccl5 -/- mice were treated with vehicle or SB225002 for 1 hour, followed by injection with α-Galcer. ( C ) Macroscopic view of liver after SB225002 treatment. ( D ) Representative H&E staining of liver tissue was shown. ( E ) Graphs represent mean score of inflammatory foci and necrotic area for each group of mice. ( F ) Serum ALT levels were measured. ( G ) Serum AST levels were measured. * P < .05, ** P < .01.

Article Snippet: Antibodies used in immunohistochemistry staining included MPO (cat number: 901-023-100510; Biocare Medical, Concord, CA) and CXCL1 (cat number: BS-10234R; Bioss, Beijing, China).

Techniques: Injection, Staining

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: Cx43 is required for TNF-α-evoked and basal release of CXCL1 in astrocyte cultures. (A and B) CXCL1 release in astrocytes following TNF-α stimulation (10 ng/ml, 60 min). Note the TNF-α-induced CXCL1 release is suppressed by pretreatment (60 min) of CBX (20 and 100 µM, A) and Gap26 and Gap27 (100 µM, B) but not by the inhibitors of pannexin hemichannels probenecid (Prob, 500 µM, A) and PANX1 mimetic peptide 10Panx1 (100 µM, A) and the scrambled peptide (Gap27 scrambled, 100 µM, B). *P < 0.05, compared with control; #P < 0.05, compared with TNF-α. (C) Inhibition of basal release of CXCL1 by CBX in astrocytes. *P < 0.05, compared with control. (D) Evoked expression (content) of CXCL1 in astrocytes following TNF-α stimulation (10 ng/ml, 60 min). Note the TNF-α-induced CXCL1 expression is not suppressed by pretreatment (60 min) of CBX (20 and 100 µM) and inhibitors of pannexin hemichannels probenecid (Prob, 500 µM) and PANX1 mimetic peptide 10Panx1 (100 µM). In contrast, a high dose of CBX (100 µM) increases CXCL1 expression. *P < 0.05, compared with control; #P < 0.05, compared with TNF-α. (E) Effects of CBX on the basal expression (content) of CXCL1 in astrocytes. All data are mean ± SEM. n = 8 cultures/group. The differences between groups were analysed by ANOVA followed by Newman–Keuls test.

Article Snippet: We also purchased SB 203580 from Calbiochem, SB 225002 from Tocris, CXCL1 neutralizing antibody from Boster, and normal Rabbit IgG from Santa Cruz.

Techniques: Control, Inhibition, Expressing

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: Spinal injection of TNF-α-activated astrocytes induces mechanical allodynia via Cx43-mediated CXCL1 release. (A) Intrathecal injection of TNF-α-activated astrocytes elicited persistent mechanical allodynia for >48 h. Note this allodynia is reduced by pretreatment of astrocytes with Cx43 small interfering RNA (1 µg/ml, 18 h). *P < 0.05, compared with TNF-α or TNF-α + non-targeting control small interfering RNA treated group; n = 6 mice/group. (B) ELISA analysis shows increased CXCL1 release in the CSF at 3 h after the intrathecal injection of TNF-α-activated astrocytes. *P < 0.05, compared with vehcile group; #P < 0.05, compared with non-activated astrocytes; n = 4 mice/group. (C) Intrathecal injection of a CXCL1 neutralizing antibody (4 µg) transiently and partially reversed mechanical allodynia, induced by TNF-α-treated astrocytes. *P < 0.05, compared with control IgG group; n = 6 mice/group. (D) Intrathecal injection of the CXCR2 antagonist SB225002 (20 µg = 57 nmol) transiently and partially reversed mechanical allodynia, induced by TNF-α-activated astrocytes. *P < 0.05, compared with vehicle (PBS); n = 5–6 mice/group. All data are mean ± SEM. The differences between groups were analysed by ANOVA followed by Newman–Keuls test.

Article Snippet: We also purchased SB 203580 from Calbiochem, SB 225002 from Tocris, CXCL1 neutralizing antibody from Boster, and normal Rabbit IgG from Santa Cruz.

Techniques: Injection, Small Interfering RNA, Control, Enzyme-linked Immunosorbent Assay

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: CXCL1, upregulated in spinal cord astrocytes after nerve injury, enhances excitatory synaptic transmission in spinal cord neurons and maintains neuropathic pain via CXCR2. (A) Western blotting shows CXCL1 upregulation in the spinal cord dorsal horn 21 days after CCI. Right, quantification of Cx43 levels in the dorsal horn. The western blot results are presented as a fold of sham control. *P < 0.05, compared to sham control, Student’s t-test, n = 4 mice/group. (B) Intrathecal injection of SB 225002 (20 µg), 21 days after CCI, reduced CCI-induced mechanical allodynia in the late phase. *P < 0.05, compared with vehicle (saline), Student’s t-test, n = 6 mice/group. (C) Double immunostaining of CXCL1 and GFAP in the dorsal horn 21 days after CCI. Note CXCL1 is primarily colocalized with GFAP. Arrows indicate doubled-labelled cells. Scale bar = 50 µm. (D and E) CXCL1 superfusion (100 ng/ml) increases spontaneous EPSC frequency (revealed by patch clamp recordings) in lamina IIo neurons of spinal cord slices. (E) Spontaneous EPSC frequency. *P < 0.05, Student’s t-test, n = 5 neurons/group. (F and G) CCI (21 d) increases spontaneous EPSC frequency, which is reversed by the CXCR2 antagonist SB225002 (1 µM). *P < 0.05, Student’s t-test, n = 5 neurons/group.

Article Snippet: We also purchased SB 203580 from Calbiochem, SB 225002 from Tocris, CXCL1 neutralizing antibody from Boster, and normal Rabbit IgG from Santa Cruz.

Techniques: Transmission Assay, Western Blot, Control, Injection, Saline, Double Immunostaining, Patch Clamp

Journal: Brain

Article Title: Connexin-43 induces chemokine release from spinal cord astrocytes to maintain late-phase neuropathic pain in mice

doi: 10.1093/brain/awu140

Figure Lengend Snippet: Schematic of working hypothesis for astrocytic Cx43-mediated late-phase neuropathic pain. CCI induces a persistent upregulation of Cx43 in spinal cord astrocytes. Cx43 expression and activity is also upregulated by TNF-α, secreted from microglia. Upregulation of Cx43 hemichannel activities results in CXCL1 release. Astrocytic CXCL1 secretion activates CXCR2 on neurons (central terminals of primary sensory neurons and spinal cord neurons), leading to enhanced excitatory synaptic transmission in nociceptive neurons (e.g. lamina IIo excitatory interneurons) and sustained neuropathic pain in the late-phase. Additionally, CXCL1 can also be secreted from intact or injured primary afferents in the spinal cord especially in the early phase of CCI.

Article Snippet: We also purchased SB 203580 from Calbiochem, SB 225002 from Tocris, CXCL1 neutralizing antibody from Boster, and normal Rabbit IgG from Santa Cruz.

Techniques: Expressing, Activity Assay, Transmission Assay

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: Clinical background and studied mechanism. (A) Global prevalence of hemodialysis (HD) and peritoneal dialysis (PD) (top left panel): hemodialysis is the most common method of RRT, while PD is thought to be underutilized by only ~11% of patients ( , ). (B) Visualization of the PD principle (top central panel): Dialysis fluid is infused into the peritoneal cavity through a catheter. The fluid absorbs toxic waste products and excess water from blood vessels of the peritoneum and then is drained into the effluent bag. (C) Medical need (top right panel): The long-term efficacy of PD is hampered by loss of the ultrafiltration capacity of the peritoneal membrane due to detrimental remodelling and angiogenesis. (D) Cellular and molecular mechanism underlying peritoneal angiogenesis studied in this manuscript (lower panel): The cytokine interleukin 17 (IL-17) acts on peritoneal mesothelial cells and activates the nuclear transcription factor SP1, which leads to CXCL1 promoter activation, mRNA production, protein synthesis and release into the extracellular space. The released CXCL1 is a potent angiogenic stimulus, and the amount of CXCL1 in the peritoneal membrane correlates with the density of peritoneal microvessels.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Membrane, Activation Assay

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: List of antibodies used in this study.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Blocking Assay, Plasmid Preparation

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: CXCL1 promotes tube formation by microvascular endothelial cells. (A) Representative microscopic images of HMECs (magnification 100x) embedded in Matrigel and treated for 16 hours with recombinant human CXCL1 at doses indicated; and (B) Quantification of the total segment length, and the number of junctions and meshes in CXCL1-treated HMECs (n=4, ANOVA) and an exemplary analysis of the parameters characterizing the endothelial network using the Angiogenesis Analyzer on ImageJ software. *P < 0.05 and **P < 0.01.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Recombinant, Software

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: Correlation between peritoneal CXCL1 expression and CD31 + microvessels and CD45+ leukocyte infiltration in PD patients and CKD5 individuals. Peritoneal histology analysis comparing peritoneal dialysis (PD; n=43) patients and chronic kidney disease stage 5 (CKD5; n=13) patients before initiation of PD for expression of CXCL1, CD31, CD45 and IL-17: (A, B) Representative immunostaining for (A) CXCL1 (insets focusing on the mesothelium, magnification 20× or 40× in insets) and (B) CD31 + microvessels and (C) Spearman correlation between peritoneal CXCL1 staining and density of CD31-positive microvessels; and (D, E) Representative immunostaining for (D) CD45 and (E) IL-17 and (F) Spearman correlation between peritoneal CXCL1 staining and number of CD45-positive leukocytes; and (G, H, I) Violin plots with quantification of obtained results for CXCL1, CD31, and CD45, depicting the individual measurements, medians and quartiles.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Expressing, Immunostaining, Staining

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: Correlation between the abundance of peritoneal CXCL1 staining and selected clinical parameters in PD patients (n=43).

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Staining

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: Conditioned medium from Il-17-stimulated mesothelial cells promotes tube formation by microvascular endothelial cells in an IL-17-dose dependent manner. Conditioned medium (CM) was collected from HPMCs treated with IL-17 for 24 hours and then added (10% v/v) to HMECs in Matrigel. After another 16 hours, the HMEC network was analysed; (A) Quantification of total segment length in HMECs exposed to CM from HPMCs stimulated with IL-17 at the indicated doses. In control, HMECs were incubated with CM derived from HPMCs not exposed to IL-17 (n=3, ANOVA). Representative images of thus treated HMECs are presented; (B) CXCL1 mRNA levels (fold change) and cytokine levels (pg/µg of cell protein) in HPMCs stimulated with IL-17 (100 ng/ml) for 24 hours in the presence of either IL-17-blocking antibody or irrelevant (irr.) control antibody at the same dose of 1 µg/ml (n=6, ANOVA); and (C) CM generated as in B was added (10% v/v) to HMECs for 16 hours and total segment lengths were assessed. In control, HMECs were treated with CM from unstimulated HPMCs (n=4-8, ANOVA vs. cells exposed to CM from HPMCs treated with IL-17 alone). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; ns, not significant.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Incubation, Derivative Assay, Blocking Assay, Generated

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: CXCL1 is a mediator of angiogenesis in mesothelial cell conditioned medium stimulated with either recombinant IL17 or IL-17-containing peritoneal dialysate. (A) HMECs were exposed for 16 hours to CM from IL-17-stimulated HPMCs in the presence of either CXCL1-neutralizing antibody or irrelevant (irr.) control IgG at the same dose (1 µg/ml), and analysed for total segment length. In control, HMECs were incubated with CM derived from HPMCs not exposed to IL-17 (n=4, ANOVA); and (B, C) HPMCs were transiently transfected with 10 nM CXCL1 siRNA, SP1 siRNA or scrambled (scramb.) control siRNA and then stimulated with IL-17 (100 ng/ml) for 24 hours and assessed for CXCL1 mRNA or protein expression (fold change or pg/ug of protein, respectively; n=6, ANOVA); and (D) CM generated as generated in (B, C) was added (10% v/v) to HMECs for 16 hours and total segment lengths were assessed. In control, HMECs were treated with CM from unstimulated HPMCs (n=4-8, ANOVA); and (E, F) CXCL1 mRNA and protein levels in HPMCs incubated for 24 hours with PD effluent (PDE, 25% v/v) from a patient with acute peritonitis. HPMCs were either transiently transfected with 10 nM siRNAs ( CXCL1 , SP1 , scrambled control) or exposed to PDE in the presence of anti-IL-17 antibody or control IgG (1 µg/ml); and (G) CM generated as in (E, F) was added (10% v/v) to HMECs for 16 hours and total segment lengths were assessed. Data are presented as the mean (± SD) fold change from HMECs exposed to CM from HPMCs treated with PDE alone (n=4-8, ANOVA). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Recombinant, Incubation, Derivative Assay, Transfection, Expressing, Generated

Journal: Frontiers in Immunology

Article Title: Angiogenic Role of Mesothelium-Derived Chemokine CXCL1 During Unfavorable Peritoneal Tissue Remodeling in Patients Receiving Peritoneal Dialysis as Renal Replacement Therapy

doi: 10.3389/fimmu.2022.821681

Figure Lengend Snippet: Spiking of recombinant IL-17 into peritoneal dialysis effluent promotes CXCL1 induction in a dose-dependent manner. PDEs from stable non-infected PD patients (n=3) were spiked with increasing doses of IL-17 (1, 10, 100 ng/ml) and applied to HPMC cultures with readout of CXCL1 protein production (pg/µg of protein) 24 hours later. ***P < 0.001, ****P < 0.0001; ns, not significant.

Article Snippet: CXCL1 , Polyclonal goat IgG , R&D Systems, Bio-Techne, Wiesbaden, Germany; (#BAF275) , Blocking , 1 µg/mL.

Techniques: Recombinant, Infection

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: Expression of CXCL1 in HCC tissue samples and HCC cell lines. (A) Determining the mRNA expression of CXCL1 in 48 cases of HCC tissues and corresponding nontumor tissues. (B) CXCL1 mRNA levels was measured by qRT-PCR as indicated HCC cell lines. (C) The protein expression levels of CXCL1 were determined by Western blot analysis in HCC cell lines. GAPDH was used as a loading control *P < 0.05, **P < 0.01.

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: Representative immunohistochemical staining of CXCL1 in HCC tissue. Representative image showed strong staining of CXCL1 (A) , moderate staining of CXCL1 (B) , nearly negative staining of CXCL1 (C) in HCC samples. All images were captured at 100× magnification.

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining, Negative Staining

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: Correlation between CXCL1 expression and clinicopathologic parameters of HCC

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: Kaplan-Meier analysis of HCC survival rates in relation to CXCL1 protein expression. Overall survival rates of patients with high (n = 33) and low (n = 15) CXCL1 expression.

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: The effect of CXCL1 on cell proliferation and invasion in 7721 and 7402 cell lines. (A) CXCL1 and p-P65 expression in the control and CXCL1-transfected cells was determined by Western blotting. GAPDH was used as a loading control. (B and C) Cell proliferation of CXCL1 was assessed in 7721 and 7402 cells by MTT at the indicated time. (D) Representative images of cells with or without CXCL1 transfection that had migrated into the lower chamber were shown. Quantitative analysis of cell invasion was assessed by invasion assay.

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Expressing, Control, Transfection, Western Blot, Invasion Assay

Journal: Cancer Cell International

Article Title: Overexpression of Chemokine (C-X-C) ligand 1 (CXCL1) associated with tumor progression and poor prognosis in hepatocellular carcinoma

doi: 10.1186/s12935-014-0086-8

Figure Lengend Snippet: P65 involved in CXCL1-induced cancer cell invasion. (A) Effect of NF-κB inhibitors on CXCL1-induced the protein expression of p-P65 in 7721 and 7402 transfected with CXCL1. (B) HCC cells 7721 and 7402 transfected with CXCL1 were treated with or without PDTC (100 μm) for 6 hours and showed reduced invasion activity compared with the negative control.

Article Snippet: After blocking with 5% nonfat milk in TBS + 0.05% TweenTM 20 for 1 hour at room temperature, membranes were incubated with an Rabbit Anti-CXCL1 antibody (Proteintech, Chicago, IL, USA), Rabbit Anti-Phospho-NF-κB p65 (Ser536) antibody, Rabbit Anti-NF-κB P65 antibody (Cell signaling Technology, Beverly, MA, USA), or Anti-GAPDH (Bioworld, Nanjing, China) overnight at 4°C.

Techniques: Expressing, Transfection, Activity Assay, Negative Control