recombinant human cxcl5  (PeproTech)

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: bioRxiv

Article Title: Tirzepatide reduces diet-induced senescence and NETosis-mediated liver fibrosis in mice

doi: 10.1101/2025.05.27.656284

Figure Lengend Snippet: a,b, Bulk RNA-seq was performed on the liver of mice fed CD or HFD for 10 weeks ( n = 3 per group). Top 10 enriched cytokine and chemokine-related GO terms ( a ) and heatmap of top 5 cytokine/chemokine DEGs based on fold change ( b ) in the liver of mice on HFD compared to mice on CD for 10 weeks. padj, adjusted p-value. Log2FC, log2FoldChange. c, Images of immunofluorescence staining of CXCL5 and Ly6G on the liver of mice fed HFD or CD for 10 weeks. DIC channel showed SBB-stained lipofuscin (black patches). Scale bar, 20 µm. d,e, dHL-60 were incubated with 50 ng/mL CXCL5 for 24 hours and stimulated with 4 µM ionomycin for 4 hours ( d ) or 1 hour ( e ). d, Representative images of NETs from dHL-60 (top) and quantification of NETs (%) (below). Yellow arrows indicate examples of NETs. n = 5 per group. Scale bar, 100 µm. e, Representative images of H4Cit high cells from dHL-60 (top) and quantification of H4Cit high cells (%) (bottom). n = 3 per group. Scale bar, 100 µm. f, dHL-60 were incubated with 50 ng/mL CXCL5 for 24 hours and stimulated with 50 µM oleic acid (OA) for 4 hours. Representative images of NETs (top) and quantification of NETs (%) (bottom). Yellow arrows indicate examples of NETs. n = 5 per group. Scale bar, 100 µm. US, unstimulated. Iono, ionomycin. Statistical comparisons were made using two-tailed unpaired t-test. * P < 0.05, ** P < 0.01 and *** P < 0.001. Data are mean ± s.e.m.

Article Snippet: To examine the effect of CXCL5 on NETosis, HL-60 were differentiated with DMSO for 6 days, and then exposed to 50 ng/mL recombinant human CXCL5 (PeproTech, 300-22) for 24 hours.

Techniques: RNA Sequencing, Immunofluorescence, Staining, Incubation, Two Tailed Test

Journal: bioRxiv

Article Title: Tirzepatide reduces diet-induced senescence and NETosis-mediated liver fibrosis in mice

doi: 10.1101/2025.05.27.656284

Figure Lengend Snippet: a-g, Male Padi4 fl/fl (Control) and Vav1-Cre Padi4 fl/fl (KO) fed CD or HFD for 10 weeks ( a ). b, Single-plane confocal images of neutrophils and NETs in the liver of control and KO mice fed HFD for 10 weeks. DIC channel showed SBB-stained lipofuscin (black patches). Scale bar, 5 µm. c, Representative images of liver interstitial fibrosis revealed by Masson’s trichrome (left) and quantification of area of interstitial fibrosis (%) (right) in the liver sections. Left to right on the bar chart, n = 5, 3, 6, 8. Scale bar, 50 µm. d, Representative images of lipofuscin stained by SBB (left) and quantification of area of SBB (%) (right) in the liver. Left to right on the bar chart, n = 5, 4, 6, 8. Scale bar, 50 µm. e, Representative Western blots (left) and quantification (right) of CXCL5 levels in the liver. Ribosomal protein lateral stalk subunit P0 (RPLP0) served as the loading control. Left to right on the bar chart, n = 4, 3, 6, 4. f, Representative images of H&E-stained liver sections from control and KO mice on HFD for 10 weeks, showing hepatocyte ballooning (green arrows), steatosis (black arrows) and immune cell infiltration (yellow squares). Scale bar, 50 µm. g, Representative images of ORO-stained liver sections (left) and quantification of area of ORO (%) (right). Left to right on the bar chart, n = 5, 4, 5, 6. Scale bar, 50 µm. Statistical comparisons were made using two-tailed unpaired t-test ( c,d,g ) or Mann-Whitney test ( e ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Data are mean ± s.e.m.

Article Snippet: To examine the effect of CXCL5 on NETosis, HL-60 were differentiated with DMSO for 6 days, and then exposed to 50 ng/mL recombinant human CXCL5 (PeproTech, 300-22) for 24 hours.

Techniques: Control, Staining, Western Blot, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Tirzepatide reduces diet-induced senescence and NETosis-mediated liver fibrosis in mice

doi: 10.1101/2025.05.27.656284

Figure Lengend Snippet: a-g, Male C57BL/6J mice were fed CD or HFD with concurrent treatment of TZP or vehicle for 10 weeks ( a ). b, Representative images of liver interstitial fibrosis revealed by Masson’s trichrome (left) and quantification of area of interstitial fibrosis (%) (middle) on FFPE liver sections. Level of liver fibrosis assessed by hydroxyproline content (µg per g liver) (right). For quantification of area of interstitial fibrosis (%), left to right on the bar chart, n = 9, 9, 10, 9. For hydroxyproline assay, left to right on the bar chart, n = 15, 13, 13, 15. Scale bar, 50 µm. c, Representative images of lipofuscin stained by SBB (left) and quantification of area of SBB (%) (right). Left to right on the bar chart, n = 9, 10, 10, 9. Scale bar, 50 µm. d, Single-plane confocal images of hepatocytes (ASGR1 + ) (left) and quantification of lipofuscin-laden hepatocytes (%) (right) in the liver sections. n = 5 per group. Scale bar, 20 µm. e, Representative Western blots (top) and quantification (bottom) of CXCL5 levels in the liver. RPLP0 served as the loading control. Left to right on the bar chart, n = 6, 6, 6, 8. f, Immunofluorescence staining for neutrophil recruitment (Ly6G + cells) in the liver (left) and quantification of neutrophils in 12 high-power fields (right). Left to right on the bar chart, n = 5, 5, 7, 7. Scale bar, 50 µm. Statistical comparisons were made using two-tailed unpaired t-test ( b-middle,c,d,e,f ) or Mann-Whitney test ( b-right ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant. Data are mean ± s.e.m. g, Single-plane confocal images of neutrophils and NETs in the liver sections from HFD-fed mice treated with vehicle or TZP. Top, yellow arrows indicate NETs; bottom, non-NETing neutrophils. Regions outlined by white dotted boxes are magnified on the immediate right. Scale bars, 5 µm.

Article Snippet: To examine the effect of CXCL5 on NETosis, HL-60 were differentiated with DMSO for 6 days, and then exposed to 50 ng/mL recombinant human CXCL5 (PeproTech, 300-22) for 24 hours.

Techniques: Hydroxyproline Assay, Staining, Western Blot, Control, Immunofluorescence, Two Tailed Test, MANN-WHITNEY

Journal: bioRxiv

Article Title: Tirzepatide reduces diet-induced senescence and NETosis-mediated liver fibrosis in mice

doi: 10.1101/2025.05.27.656284

Figure Lengend Snippet: a-k, Male C57BL/6J mice were fed CD or HFD for 7 weeks, followed by treatment with TZP or vehicle for the subsequent 10 weeks along with the respective diets ( a ), and bulk RNA-seq was performed on the liver tissue from these mice ( n = 3 per group) ( g-k ). b, Representative images of liver interstitial fibrosis revealed by Masson’s trichrome (left) and quantification of area of interstitial fibrosis (%) (middle) on the FFPE liver sections. Level of liver fibrosis assessed by hydroxyproline content (µg per g liver) (right). For quantification of area of interstitial fibrosis (%), left to right on the bar chart, n = 8, 6, 8, 7. For hydroxyproline assay, left to right on the bar chart, n = 3, 5, 5, 5. Scale bar, 50 µm. c, Representative images of lipofuscin stained by SBB (left) and quantification of area of SBB (%) (right) of the liver sections. Left to right on the bar chart, n = 7, 7, 8, 8. Scale bar, 50 µm. d, Representative Western blots (top) and quantification of CXCL5 levels (bottom) in the liver. RPLP0 served as the loading control. Left to right on the bar chart, n = 4, 4, 5, 5. e, Immunofluorescence staining for neutrophil recruitment (Ly6G + cells) in the liver (left) and quantification of neutrophils in 12 high-power fields (right). Left to right on the bar chart, n = 5, 5, 8, 8. Scale bar, 50 µm. Statistical comparisons were made using two-tailed unpaired t-test ( b,d,e ) or Mann-Whitney test ( c ). * P < 0.05, ** P < 0.01 and *** P < 0.001. Data are mean ± s.e.m. f, Single-plane confocal images of neutrophils and NETs in the liver of HFD-fed mice treated with vehicle or TZP. Top, yellow arrows indicate NETs; bottom, non-NETing neutrophils. Regions outlined by the white dotted boxes are magnified on the immediate right. Scale bars, 5 µm (left) and 2 µm (right). g, Principal component analysis of the bulk RNA-seq data from the liver of mice with different treatments. h, Top 20 enriched GO terms based on adjusted p-value in the liver of mice fed HFD with and without TZP treatment. i-k, Heatmap of DEGs in fibrosis-related pathways (extracellular matrix organization, transforming growth factor beta receptor signaling pathway, fibroblast proliferation, collagen biosynthetic process, wound healing) ( i ), lipofuscin-related pathways (autophagosome-lysosome fusion, response to oxidative stress, cellular response to oxidative stress, intrinsic apoptotic signaling pathway in response to oxidative stress, lipid oxidation, macroautophagy, TOR signaling) ( j ), and neutrophil-related pathways (neutrophil activation, neutrophil chemotaxis, neutrophil migration) ( k ).

Article Snippet: To examine the effect of CXCL5 on NETosis, HL-60 were differentiated with DMSO for 6 days, and then exposed to 50 ng/mL recombinant human CXCL5 (PeproTech, 300-22) for 24 hours.

Techniques: RNA Sequencing, Hydroxyproline Assay, Staining, Western Blot, Control, Immunofluorescence, Two Tailed Test, MANN-WHITNEY, Activation Assay, Chemotaxis Assay, Migration

Journal: bioRxiv

Article Title: Tirzepatide reduces diet-induced senescence and NETosis-mediated liver fibrosis in mice

doi: 10.1101/2025.05.27.656284

Figure Lengend Snippet: Choline-deficient L-amino acid-defined high-fat diet (CDAHFD) induces lipofuscin deposition in the lysosomes of hepatocytes. Lipofuscin-containing cells produce higher levels of CXCL5, which in turn enhances NET formation, eventually leading to liver fibrosis. Intercepting NETosis through direct inhibition of peptidylarginine deiminase 4 (PAD4) or through decreasing hepatic lipofuscin load by TZP treatment both effectively reduce liver fibrosis. TZP treatment provides additional therapeutic benefits by lessening hepatic cellular senescence. The illustration was created with BioRender.com.

Article Snippet: To examine the effect of CXCL5 on NETosis, HL-60 were differentiated with DMSO for 6 days, and then exposed to 50 ng/mL recombinant human CXCL5 (PeproTech, 300-22) for 24 hours.

Techniques: Inhibition