Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17 enhances the net angiogenic activity and in vivo growth of human non-small cell lung cancer in SCID mice through promoting CXCR-2-dependent angiogenesis.

doi: 10.4049/jimmunol.175.9.6177

Figure Lengend Snippet: FIGURE 1. IL-17R expression on human NSCLC lines. Three different human NSCLC lines, Sq-19, A549, and LK-87, were incubated with mouse anti-human IL-17R mAb (solid line) or irrelevant mouse IgG1 (dotted line), followed by PE-conjugated goat anti-mouse IgG1 Ab. The expression of IL-17R on the surface of NSCLC cells was analyzed by flow cytometry.

Article Snippet: Cells (1 106), washed twice with PBS containing 2% FCS and 0.1% NaN3, were incubated with mouse anti-human IL-17R mAb or with irrelevant normal mouse IgG1 (R&D Systems) at 4°C for 60 min. After washing, the cells were incubated with PE-conjugated goat anti-mouse IgG1 Ab (Caltag Laboratories) at 4°C for 30 min.

Techniques: Expressing, Incubation, Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: IL-17 enhances the net angiogenic activity and in vivo growth of human non-small cell lung cancer in SCID mice through promoting CXCR-2-dependent angiogenesis.

doi: 10.4049/jimmunol.175.9.6177

Figure Lengend Snippet: FIGURE 9. IL-17 and IL-17R expression and vascularity in human NSCLC tissues. A, Total cellular RNA was extracted from human NSCLC tissues. Five micrograms of total RNA were applied for the synthesis of cDNAs. IL-17 mRNA expression was detected in five of seven NSCLC

Article Snippet: Cells (1 106), washed twice with PBS containing 2% FCS and 0.1% NaN3, were incubated with mouse anti-human IL-17R mAb or with irrelevant normal mouse IgG1 (R&D Systems) at 4°C for 60 min. After washing, the cells were incubated with PE-conjugated goat anti-mouse IgG1 Ab (Caltag Laboratories) at 4°C for 30 min.

Techniques: Expressing

Journal: Cell reports

Article Title: IL-17/CXCL5 signaling within the oligovascular niche mediates human and mouse white matter injury

doi: 10.1016/j.celrep.2022.111848

Figure Lengend Snippet:

Article Snippet: The following primary antibodies were used: mouse anti-NF200 (1:200, Sigma), rabbit anti-MBP (1:500, Calbiochem), goat anti-PDGFRα (1:500; Neuromics), mouse anti-HA (1:1000, Biolegend), rabbit-Gst-π (1:1000, Millipore), rabbit anti-IL-17Rb (1:500, Santa Cruz Biotech), rat anti-CXCL5 (1:250, R&D) in PBS containing 5% goat or donkey serum and 0.3% Triton-X 100 (Sigma) overnight at 4°C.

Techniques: Blocking Assay, Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Luminex, Hybridization, Plasmid Preparation, Biomarker Discovery, Control, Software

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: The sequences of primer pairs used in qPCR.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques:

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: IL-17RB but not IL-17RA is involved in the skin wound healing of patients with DFU. (A–E) Relative mRNA expression of IL-25, IL-17RB, IL-17RA, IL-9 and VEGF in the wound edge skin from diabetic patients with DFU (n = 11) and normoglycemic individuals (n = 7), respectively. (F, G) Representative immunofluorescent staining for IL-17RB (green) and DAPI (blue) in the wound edge skin from diabetic patients with DFU and normoglycemic individuals. Scale bars, 20 μm. Quantification of the IL-17RB protein in the skin is shown in the histogram in G (n = 3, three views for each sample). * P < 0.05, ** P < 0.01, *** P < 0.001. ns, no significance.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: Expressing, Staining

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: Downregulation of IL-25-mediated-IL-17RB signaling contributes to the delay of wound healing in diabetic mice. 7-day wound biopsies within 2 mm of a wound edge were obtained from dorsal skin of control (CON) and diabetic (DM) mice. (A) Images of representative wounds on day 0 and day 11 after injury. (B) Quantification of time of wound closure (n = 5). (C) Representative images of Masson trichrome staining of dorsal skin sections from control and diabetic mice. Scale bars, 200 μm (left), 25 μm (right). (D) Quantification of collagen deposition in different groups (n = 3, three views for each sample). (E) Angiogenesis analysis by immunohistochemistry staining of CD31 in dorsal skin sections from control and diabetic mice. Scale bars, 25 μm. (F) Quantification of the vessel numbers is displayed in the graph (n = 3). (G, H) Relative mRNA expression of IL-25 and IL-17RB in the wound skin from control and diabetic mice (n = 3). (I, J) Representative immunofluorescent staining for IL-17RB (red) and DAPI (blue) in skin sections of wound edge from control and diabetic mice. Scale bars, 20 μm. Quantification of the IL-17RB protein expression in the mice skin is shown in the histogram in J (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: Control, Staining, Immunohistochemistry, Expressing

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: IL-25-upregulated-IL-17RB facilitates wound healing in diabetic mice by rescuing β-catenin expression. (A) Representative immunofluorescent staining for IL-17RB (red) and DAPI (blue) in wound edge skin (day 7 post injury) from groups of control (CON), diabetic (DM) and diabetic with rmIL-25 injection mice (DM + rmIL-25). Scale bars, 20 μm. (B) Quantification of IL-17RB in A is shown (n = 3). (C) Representative western blotting image of β-catenin and IL-17RB in wound skin (day 7 post injury) from groups of CON, DM and DM + rmIL-25. (D) Quantification analysis of β-catenin and IL-17RB protein in C is presented (n = 3). (E, F) Protein expression level of β-catenin (green) in wound skin (day 7 post injury) from group of CON, DM and DM + rmIL-25 was evaluated by immunofluorescent staining (E) and the quantification analysis was shown (n = 3) (F) . Scale bars, 20 μm. (G) Representative immunofluorescent staining for β-catenin (green) and DAPI (blue) in wound edge skin from diabetic patients with DFU and normoglycemic individuals. Scale bars, 20 μm. (H) Quantification analysis of the β-catenin protein expression from G (n = 3). (I) β-catenin mRNA expression in the wound edge skin from diabetic patients with DFU (n = 11) and normoglycemic individuals (n = 7) was analyzed by qPCR. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: Expressing, Staining, Control, Injection, Western Blot

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: IL-25 rescues the inactivation of Wnt/β-catenin pathway in vitro . (A, B) HUVECs were stimulated with increasing concentrations of rhIL-25 for 48 h under high glucose conditions (HG). β-catenin and cyclin D1 mRNA from untreated HUVEC and rhIL-25-treated HUVECs under HG conditions were analyzed by qPCR analysis (n = 3). (C) IL-17RB mRNA expression in HUVECs stimulated with 400 ng/mL rhIL-25 under HG conditions for 48 h (n = 3). (D, E) Western blotting analysis of β-catenin and IL-17RB and their quantification (n = 3). (F–I) Representative immunofluorescent staining images and quantification of β-catenin (F, G) and IL-17RB (H, I) in HUVECs exposed to high glucose and high glucose with rhIL-25 conditions for 48 h (n = 3). Scale bars, 60 μm. * and # P < 0.05, ** P < 0.01, *** P < 0.001 ( # compared with CON group, *compared with HG group). ns, no significance.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: In Vitro, Expressing, Western Blot, Staining

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: Endogenous overexpression of IL-25 activates Wnt/β-catenin pathway. HUVECs that stably overexpress IL-25 (Lenti-IL-25-GFP-HUVECs) were constructed by using a lentiviral expression vector of human interleukin 25 with the unfused CopGFP protein at the C-terminus (Lenti-IL-25-GFP). The overexpression of IL-25 was verified by q-PCR and immunofluorescent staining assays. (A) Relative IL-25 mRNA expression in wild type (WT) and Lenti-IL-25-GFP-HUVECs (n = 3). (B) Immunofluorescent staining images of IL-25 (red), GFP (green) and DAPI (blue) in wild type (WT) and Lenti-IL-25-GFP-HUVECs. Scale bars, 20 μm. (C, D) Western blotting analysis of β-catenin and IL-17RB in WT and Lenti-IL-25-GFP-HUVECs and their quantification under normal glucose conditions (n = 3). (E, F) Western blotting analysis of β-catenin and IL-17RB in WT and Lenti-IL-25-GFP-HUVECs after exposure to high glucose conditions for 24 and 48 h and their quantification (n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001. ns, no significance.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: Over Expression, Stable Transfection, Construct, Expressing, Plasmid Preparation, Staining, Western Blot

Journal: Frontiers in Immunology

Article Title: Interleukin-25-Mediated-IL-17RB Upregulation Promotes Cutaneous Wound Healing in Diabetic Mice by Improving Endothelial Cell Functions

doi: 10.3389/fimmu.2022.809755

Figure Lengend Snippet: IL-25 induces AKT and ERK 1/2 phosphorylation in HUVECs under high glucose conditions. (A, B) HUVECs were cultured in 6-well plate. After grown to 80%, cells were treated with high glucose medium in the presence of a gradient of rhIL-25. Proteins were collected for western blotting analysis after culturing for another 48 h. Representative western blotting images of p-AKT, p-ERK 1/2 and IL-17RB (A) and their quantification (B) (n = 3) were shown. (C, D) WT-HUVECs and Lenti-IL-25-GFP-HUVECs were cultured in normal glucose conditions and proteins were collected for western blotting analysis. Representative western blotting images of p-AKT and p-ERK 1/2 (C) and their quantification is displayed in the graph (D) (n = 3). (E, F) WT-HUVECs and Lenti-IL-25-GFP-HUVECs were treated with high glucose for 24 or 48 h and the proteins were obtained for western blotting analysis. Representative western blotting images (E) and quantification (F) of p-AKT and p-ERK 1/2 (n = 3) were shown. * and # P < 0.05, ** P < 0.01, *** and ### P < 0.001 ( # compared with CON group, *compared with HG group). ns, no significance.

Article Snippet: Goat anti-IL-17RB antibody (Cat#: AF1040) was obtained from R&D systems.

Techniques: Phospho-proteomics, Cell Culture, Western Blot