Journal: Nature Communications

Article Title: Nlrp12 mutation causes C57BL/6J strain-specific defect in neutrophil recruitment

doi: 10.1038/ncomms13180

Figure Lengend Snippet: ( a ) WT (B6N) or Nlrp12 −/− mice were infected i.n. with 5 × 10 3 colony-forming unit of F. tularensis LVS. Three days post infection BAL were performed and cytokine levels were determined by ELISA ( n =8, WT; n =9, Nlrp12 −/−) . ( b ) BMDM from WT (B6N) and Nlrp12 −/− mice were challenged with either F. tularensis LVS, S. aureus or P. aeruginosa . Eight hours later supernatants were collected and secretion of the indicated cytokine or chemokine quantified by ELISA. ( c ) BMDM from WT (B6N) and Nlrp12 −/− mice were stimulated with LPS (50 ng ml −1 ) for the indicated amount of time; supernatants were collected and assayed for CXCL1 production by ELISA. ( d ) BMDM from WT (B6N) and Nlrp12 −/− mice were left unstimulated or stimulated for 4 h with LPS in the presence of brefeldin A and monensin; intracellular CXCL1 was then assessed by flow cytometry. ( e ) BMDM from WT (B6N) and Nlrp12 −/− mice were stimulated with LPS for the indicated amount of time; Cxcl1 expression was quantified by real-time PCR. ( f ) BMDM from WT (B6N), WT (B6J) and Nlrp12 −/− mice were stimulated with LPS for 8 h; supernatants were collected and assayed for the indicated cytokines by ELISA. ( g ) BMDM from B6N, B6J, B6N-B6J F1 and B6N-B6J F2 mice were challenged with LPS for 8 h; supernatants were collected and CXCL1 secretion assessed by ELISA. The Nlrp12 allele was sequenced for all B6N-B6J F2 mice and cohorts stratified based on their Nlrp12 genotype ( Nlrp12 B6N/B6N , Nlrp12 B6N/B6J or Nlrp12 B6J/B6J ) ( n =14, B6N; n =13, B6J; n =14, F1; n =52, F2). Pooled data from three ( b , c ) independent experiments are depicted or are representative of two ( d ) or three ( e , f ) independent experiments. ( b , c , e , f ) Data are expressed as the mean±s.e.m. * P <0.05, ** P <0.01, *** P <0.005, NS, not significant by Mann–Whitney U -test ( a , g ), Student's t -test ( b , e , f ) or two-way analysis of variance analysis ( c ).

Article Snippet: Antibody pairs for the IL-1β (MAB401 and BAF401; clone 30311 and polyclonal; R&D Systems; 8 and 4 μg ml −1 , respectively) and CXCL1 (mouse: MAB453 and BAF453; clone 48415 and polyclonal; R&D Systems; 8 and 0.4 μg ml −1 , respectively; human: MAB275, clone 20326, 4 μg ml −1 ; BAF275, polyclonal, 40 ng ml −1 ) and CCL3/ MIP-1α (Quantikine ELISA kit, Product #MMA00) ELISAs were from R&D Systems.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY

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: BMB Reports

Article Title: Regorafenib prevents the development of emphysema in a murine elastase model

doi: 10.5483/BMBRep.2023-0072

Figure Lengend Snippet: Effect of regorafenib on PPE-induced in inflammatory mediators in BAL fluid. (A) Representative membrane images of the antibody array. (B) Quantification of relative optical density for IL-1β, IL-6, CXCL1/KC, and TIMP-1 in the array membranes. (C-F) Measurement of IL-1β (C), IL-6 (D), CXCL1/KC (E), and TIMP-1 (F) levels in BAL fluid by ELISA. Error bars indicate the mean ± SEM. N = 8 per group, *P < 0.05, **P < 0.01, ****P < 0.0001, t -test.

Article Snippet: Anti-IL-6 antibody (ab208113) from abcam (Cambridge, UK), CXCL1 antibody (12335-1-AP) from Proteintech (Rosemont, IL, USA), and TIMP-1 antibody (#AF980) from R&D Systems were purchased respectively.

Techniques: Membrane, Ab Array, Enzyme-linked Immunosorbent Assay

Journal: BMB Reports

Article Title: Regorafenib prevents the development of emphysema in a murine elastase model

doi: 10.5483/BMBRep.2023-0072

Figure Lengend Snippet: Effect of regorafenib on PPE-induced expression of inflammatory mediators in lung parenchyme. (A) Experimental scheme. Mice were sensitized by treatment with regorafenib (5 mg/kg) for one day in the only PPE + Reg group. After PPE treatment, regorafenib was administered orally to mice for six days. On the seventh day, lung tissues were removed from mice, prepared in paraffin sections and stained with antibodies for inflammatory mediators. (B) Representative images of stained paraffin sections. Red arrows indicate the macrophages in inserted boxes. Scale bars indicate 50 μm. (C-E) Quantification of . IL-6 (C), CXCL1/KC (D), and TIMP-1 (E). Error bars indicate the mean ± SEM. N = 5 per group, *P < 0.05, **P < 0.01, t -test.

Article Snippet: Anti-IL-6 antibody (ab208113) from abcam (Cambridge, UK), CXCL1 antibody (12335-1-AP) from Proteintech (Rosemont, IL, USA), and TIMP-1 antibody (#AF980) from R&D Systems were purchased respectively.

Techniques: Expressing, 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: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

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: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

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: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

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: The sections were then incubated overnight at 4°C with the following primary antibodies: GFAP antibody (1:1000, mouse; Millipore Bioscience Research Reagents), Cx43 antibody (1:1000, rabbit; Sigma), NeuN antibody (1:1000, mouse; Millipore Bioscience Research Reagents), CXCL1 (1: 200, rabbit; Boster), and CXCR2 antibody (1: 200, rabbit; Boster).

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