recombinant human cxcl5 protein  (R&D Systems)

Journal: Cardiovascular diabetology

Article Title: CXCL5 suppression recovers neovascularization and accelerates wound healing in diabetes mellitus.

doi: 10.1186/s12933-023-01900-w

Figure Lengend Snippet: Fig. 2 Treatment with CXCL5 neutralizing antibody upregulated VEGF/SDF-1 expression and promoted angiogenesis in late-EPCs from non-DM subjects and HAECs under the HG conditions. The network formation and migration abilities were improved after the administration of CXCL5 mAb in EPCs from non-DM subjects (n = 3; A, B). Western blotting and statistical analyses of VEGF and SDF-1 in EPCs from non-DM subjects (n = 3; C). The network formation and migration abilities were improved after the administration of CXCL5 mAb in HAECs (n = 3; D, E). Western blotting and statistical analyses of VEGF and SDF-1 in HAECs (n = 3; F). CXCL5 C-X-C motif chemokine ligand 5, EPC endothelial progenitor cell, HG high glucose, HAEC human aortic endothelial cell, mAb,monoclonal antibody, SDF-1 stromal cell-derived factor 1, VEGF vascular endothelial growth factor. N represents the number of independent experiments on different days and in different experimental runs. The Mann–Whitney U test was used to determine statistically significant differences. *p < 0.05, **p < 0.01

Article Snippet: Some cells were treated with CXCL5 monoclonal antibody (1 or 10 μg/ mL; R&D Systems, MAB-254, Minneapolis, MN, USA) or recombinant human CXCL5 protein (1 or 10 ng/mL; R&D Systems, 254-XB, Minneapolis, MN, USA).

Techniques: Expressing, Migration, Western Blot, Derivative Assay, MANN-WHITNEY

Journal: Cardiovascular diabetology

Article Title: CXCL5 suppression recovers neovascularization and accelerates wound healing in diabetes mellitus.

doi: 10.1186/s12933-023-01900-w

Figure Lengend Snippet: Fig. 7 Summary of beneficial effects of CXCL5 suppression in diabetic vasculopathy. CXCL5 Chemokine C-X-C motif ligand 5, CXCR2 Chemokine C-X-C motif receptor 2, EPC endothelial progenitor cell, ERK extracellular signal-regulated kinase, DM diabetes mellitus, IL interleukin, SDF-1 stromal cell-derived factor 1, TNF-α tumor necrosis factor-α, VEGF vascular endothelial growth factor

Article Snippet: Some cells were treated with CXCL5 monoclonal antibody (1 or 10 μg/ mL; R&D Systems, MAB-254, Minneapolis, MN, USA) or recombinant human CXCL5 protein (1 or 10 ng/mL; R&D Systems, 254-XB, Minneapolis, MN, USA).

Techniques: Derivative Assay

Journal: Microbiology Spectrum

Article Title: Characterization of the Inflammatory Response Evoked by Bacterial Membrane Vesicles in Intestinal Cells Reveals an RIPK2-Dependent Activation by Enterotoxigenic Escherichia coli Vesicles

doi: 10.1128/spectrum.01115-23

Figure Lengend Snippet: Proinflammatory cytokine response to MVs derived from diverse intestinal bacteria in human intestinal cells. ENA-78/CXCL5 (A), GRO-alpha/CXCL1 (B), IL-8/CXCL8 (C), IP-10/CXCL10 (D), MDC/CCL22 (E) and MIP-3alpha/CCL20 (F) were detected, and levels were quantified by ELISA in supernatants of HT-29 intestinal cells incubated for 16 h with equal amounts of MVs derived from the bacterial species indicated on the x axis, respectively. MVs are sorted alphabetically by their donor species separated by Gram-negative (left) and Gram-positive (right) bacteria. Incubation with saline served as a negative control (no MVs [far right]). The corresponding basal level of each cytokine/chemokine produced by HT-29 is given by the median value of the control (no MVs) and highlighted with a horizontal black line. Data are indicated as the median ± interquartile range. (A) n = 4 for B. thetaiotaomicron , F. nucleatum and Y. enterocolitica ; n = 11 for E. cloacae and no MVs, n = 7 for enteroaggregative E. coli (EAEC) 55989, EPEC E2348/69, and L. acidophilus ; n = 8 for EAEC 042, EAEC 17-2, enteroinvasive E. coli (EIEC) EDL 1284, EIEC HN280, ETEC 1392-75, uropathogenic E. coli (UPEC) CFT073, K. oxytoca , S. flexneri , and Pediococcus acidilactici ; n = 10 for ETEC 10407 (WT) and E. coli Nissle; n = 5 for P. vulgaris ; n = 9 for Shigella sonnei and V. cholerae ; n = 6 for all other data sets. (B) n = 8 for B. fragilis , EAEC 55989, EAEC 042, EAEC 17-2, EIEC EDL 1284, EIEC HN280, ETEC 1392-75, EPEC E2348/69, P. vulgaris , S. Typhimurium, and Y. enterocolitica ; n = 12 for Bacteroides thetaiotaomicron , Bacteroides vulgatus , E. cloacae , UPEC CFT073, E. coli Nissle, and K. oxytoca ; n = 18 for ETEC 10407 (WT); n = 9 for UPEC 536; n = 14 for no MVs; n = 10 for all other data sets. (C) n = 12 for E. cloacae , K. oxytoca , S. sonnei , and L. acidophilus ; n = 47 for ETEC 10407 (WT); n = 16 for UPEC CFT073 and V. cholerae ; n = 4 for UPEC UTI89; n = 14 for E. coli Nissle; n = 28 for no MVs; n = 8 for all other data sets; (D) n = 6 for B. fragilis , UPEC CFT07, UPEC 536, K. pneumoniae , P. vulgaris , V. cholerae , and Y. enterocolitica ; n = 11 for B. vulgatus ; n = 10 for EIEC EDL 1284; n = 12 for ETEC 10407 (WT) and no MVs; n = 4 for F. nucleatum ; n = 7 for L. acidophilus ; n = 8 for all other data sets. (E) n = 12 for B. thetaiotaomicron , V. cholerae , P. acidilactici , and no MVs; n = 14 for ETEC 10407 (WT); n = 10 for E. cloacae and K. oxytoca ; n = 8 for all other data sets. (F) n = 4 for EIEC EDL 1284; n = 16 for ETEC 10407 (WT) and V. cholerae ; n = 12 for E. coli Nissle; n = 14 for no MVs; n = 8 for all other data sets.

Article Snippet: Recombinant human CXCL5, CXCL8, and CXCL10 from Biolegend served as standards (573409, 570909, and 573509).

Techniques: Derivative Assay, Bacteria, Enzyme-linked Immunosorbent Assay, Incubation, Saline, Negative Control, Produced, Control

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: Schematic representation of IL-17/CXCL5 signaling in chronically injured cerebral endothelia (A). TRAP-qPCR fold expression compared with average fpkm of top DEGs from white matter endothelia (*adjusted p < 0.05) (B). Weight-adjusted ELISA values (pg/mL) for murine CXCL5 in retro-orbital blood samples from CFD (black) and HFD (red) animals (n = 4/grp, p = 0.0355) (C). Immunofluorescence labeling for IL-17Rb (green, D) and CXCL5 (green, E) is absent in white matter vasculature of Tie2-Cre;tdTomato mice on CFD (left panels) and abundant in white matter vasculature of Tie2-Cre;tdTomato mice on HFD (right panels). Single-channel labeling for IL17Rb (bottom panels, D) and CXCL5 (bottom panels, E) show heterogeneous endothelial expression. Labeling for GLUT-1 (blue), CXCL5 (red), and PDGFRα (green) at 7 days post-stroke in animals on CFD (left) and HFD (right). Inset boxes from the peri-infarct tissue (top) masked for GLUT-1 (white) with only co-localized CXCL5 (purple) (bottom). Graph of percentage of co-localized CXCL5+/GLUT-1+ voxels (****p < 0.0001) (F). Error bars represent S.E.M. Scale bars: 50 μm (F), 20 μm (D), and 10 μm (E).

Article Snippet: Rabbit anti-human CXCL5/6 , Abcam , ab198505.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Labeling

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: Human brain microvascular endothelial cells were stimulated with IL-17 ligands A–E (250 ng/mL) and CXCL5 levels measured in conditioned media 48 h after stimulation (*p = 0.0372 by Kruskal-Wallis H test; **post-hoc comparison for IL-17B versus no ligand, adjusted p = 0.0178) (A). Phalloidin+ cellular area in O4+ OPCs grown in vitro exposed to vehicle (top panel) or recombinant CXCL5 (bottom panel) for 48 h (p < 0.0001, F = 9.82 by one-way ANOVA) (B). Approach for CXCL5 transgenic-viral gain of function in subcortical white matter of Tie2-Cre;tdTomato mice (top panel) (C). PDGFRα+ OPC (green) labeling in GFP-transduced Tie2-Cre;tdTomato mice (red, left panel) and CXCL5-GFP-transduced Tie2-Cre;tdTomato mice (right panel). Representative masked cellular profiles of PDGFRα+ cell area (bottom panels). Schematic of anti-IL-17B antibody treatment (top panel) (D). PDGFRα+ OPC (green) labeling in control IgG-treated Tie2-Cre:tdT mice (left panel) and anti-IL-17B IgG-treated Tie2-Cre:tdT mice (right panel). Representative masked cellular profiles of PDGFRα+ cell area (bottom panels). Proportion of OPCs per unit distance from vessel (0–35 μm) in each condition (total measured cell number per condition in parentheses) (E). Average distance of OPCs to vessel (***p = 0.0005, F = 6.06 by one-way ANOVA; **adjusted p = 0.0039; *adjusted p = 0.0168) (F). Average in vivo PDGFRα+ OPC cell area (**p = 0.0068, F = 7.38 by one-way ANOVA; **adjusted p = 0.002) (G). Graph of co-localized CXCL5+/GLUT-1+ voxels in anti-IL-17B IgG-treated animals (n = 4/grp; *p = 0.018) (H). Error bars represent S.E.M. Scale bars: 10 μm

Article Snippet: Rabbit anti-human CXCL5/6 , Abcam , ab198505.

Techniques: In Vitro, Recombinant, Transgenic Assay, Labeling, In Vivo

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: Plasma levels of log 10 -CXCL5 in ASPIRE cohort subjects separated by detectable plasma IL-17B (n = 32; median 1043.0 pg/mL) compared with those with undetectable plasma IL-17B (n = 99; median 515.3 pg/mL; *p < 0.0001). Plasma log 10 -CXCL5 levels in subjects with MRI-confirmed acute microvascular ischemia (IL-17B + subjects; n = 9; 978.2 pg/mL versus IL-17B− subjects; n = 24; 539.7 pg/mL) (**p = 0.0157) (A). Ordinal shift analysis of modified Fazekas scale scores from plasma IL-17B+ and IL-17B− subjects (p < 0.0001) (B). Representative immunohistochemical detection of CXCL5 in human frontal white matter vasculature in subjects with cerebrovascular pathology (C). Percentage of CXCL5+ vessel segments in peri-ventricular white matter (n = 10) (p = 0.0005). Error bars represent S.E.M. Scale bar: 10 μm

Article Snippet: Rabbit anti-human CXCL5/6 , Abcam , ab198505.

Techniques: Modification, Immunohistochemical staining

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: Rabbit anti-human CXCL5/6 , Abcam , ab198505.

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