Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 1. CXCL5 protein expression is concordant with prostate cancer progression. Shown are representative panels from a hematox- ylin and eosin–stained, high-density tissue microarray probed with antibody against CXCL5, as follows: (A) Benign glands demonstrating weak staining. (B) PCa (Gleason sum 3 + 3) demonstrating weak staining. (C) PCa (Gleason sum 4 + 4) demonstrating moderate to strong staining. (D) Hormone refractory METs demonstrating strong staining. (E) PCa demonstrating moderate to strong staining asso- ciated with stromal inflammatory component (yellow arrows point to areas of inflammation). (F) Benign glands demonstrating strongly staining luminal secretions (black arrows). Original magnifications, ×100. Panel E has been enlarged further, ×4, to illustrate the area of inflammatory infiltrate concomitant with CXCL5 protein expression. (G) Boxplot depicting median product score distributions of protein expression levels for benign glands, malignant glands from PCa, and malignant areas from METs and P values associated with the statistical evaluation of these distributions.

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Expressing, Staining, Microarray

Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 2. Nontransformed and transformed prostate epithelial cells express the CXCL5 receptor and endogenously secrete CXCL5. (A) Immunoblot analysis of protein lysates prepared from transformed PC3 and LNCaP, and nontransformed N15C6 and BPH-1 prostate epithelial cells probed with antibodies specific for the CXCL5 receptor, CXCR2, and loading control, β-actin. Pri- mary antibody concentrations used were 1:1000 for CXCR2 and 1:5000 for β-actin. (B) Protein levels (pg/ml) of CXCL5 present in media conditioned by transformed LNCaP and PC3 or nontrans- formed N15C6 or BPH-1 cells prostate epithelial cells were deter- mined by ELISA. The graph shows the pg/ml CXCL5 detected plotted on a logarithmic scale (y axis).

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Transformation Assay, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 4. CXCL5-stimulated proliferative and invasive responses. (A) N15C6 (light gray bars) or BPH-1 (dark gray bars) nontransformed prostate epithelial cells proliferated to significantly higher levels when grown for 72 hours in SF media supplemented with 10 pM CXCL5 than those grown in SF alone (*P < .001). Preincubation of the cells for 1 hour with 1 μg/ml antibody against CXCR2, the receptor for CXCL5, followed by supplementation with CXCL5 and maintenance of growth in CXCL5 + anti-CXCR2–containing media significantly ablated the proliferative response (#P < .001). In contrast, cellular growth after preincubation with an antibody against an unrelated chemokine receptor, CXCR4, followed by supplementation with CXCL5 and maintenance of growth in CXCL5 + anti-CXCR4–containing media was similar to that observed for non–pretreated cells grown in CXCL5-supplemented media and was significantly higher than that in SF alone (*P < .001). All data are shown normalized to growth in unsupplemented SF, which was set at one-fold. (B) N15C6 (LEFT) or LNCaP (RIGHT) cells were grown in SF media (untreated, UnT) or SF media supplemented with 10 pM CXCL5 for N15C6 or 100 pM CXCL5 for LNCaP (treated, T) for the times indicated. The cells were then harvested and assessed for nucleosomal DNA fragmentation. The fraction of cells exhibiting apoptosis plotted on the y axis was calculated as the difference in absorbance measured at 405 nm and at the reference wavelength of 490 nm after adjusting for background absorbance at both wavelengths. No significant differences in the fraction of cells exhibiting apoptosis were observed between treated and untreated cells at any time point, demonstrating that CXCL5 does not promote antiapoptotic responses in these cells. (C) Fifteen thousand each of N15C6 (black bars) or PC3 (gray bars) cells were plated onto Matrigel-coated membranes and were exposed to complete media or complete media supplemented with 20 nM CXCL5 for 24 hours. After 24 hours, the cells that migrated and invaded through the Matrigel were stained and counted. N15C6 cells did not demonstrate an invasive response to treatment with CXCL5. However, approximately six-fold more PC3 cells migrated through the synthetic basement membrane, Matrigel, in response to 20 nM CXCL5 compared to vehicle (control, set at one-fold) (*P < .05). PC3 cell invasion through the Matrigel in response to CXCL5 was significantly inhibited by pretreatment with 1 μg/ml blocking antibody (anti- CXCR2) (#P < .05) but not by pretreatment with nonspecific antibody (anti-CXCR4) (*P < .05).

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Staining, Membrane, Control, Blocking Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 5. CXCL5 activates MAPK signaling in nontransformed N15C6 prostate epithelial cells. Nontransformed N15C6 cells rapidly and transiently phosphorylated ERK 1/2 and STAT3 when treated with either subnanomolar (10 or 100 pM) or nanomolar (1 nM) levels of CXCL5, whereas NF-κB subunit activation was evident only after treatment with 1 nM CXCL5. Primary antibody concentrations used were 1:500 for phospho-ERK, 1:500 for phospho-65 (NF-κB), 1:1000 for phospho-STAT3, 1:1000 for total ERK, 1:1000 for total p65, and 1:2000 for total STAT3. A total of 20 μg of protein lysate was electrophoresed per well. Immunoblots are shown on the left, and corresponding densitometric evaluations of the same blots are shown on the right. Phosphorylation relative to total protein quantitated from the immunoblot is shown in the densitometric plots as phospho/total protein.

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Activation Assay, Western Blot, Phospho-proteomics

Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 6. CXCL5 activates both MAPK and PI3K signaling in transformed LNCaP prostate epithelial cells. Transformed LNCaP cells rapidly and transiently phosphorylated both ERK 1/2 and the p65 subunit of NF-κB on treatment with subnanomolar (10 or 100 pM) levels of CXCL5. Immunoblots are shown in the top panel, and corresponding densitometric evaluations of the same blots are shown in the bottom panel. Phosphorylation relative to total protein quantitated from the immunoblot is shown in the densitometric plots as phospho/total protein. A total of 100 μg of protein lysate was electrophoresed per well. Primary antibody concentrations used were as described for Figure 5.

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Transformation Assay, Western Blot, Phospho-proteomics

Journal: Neoplasia (New York, N.Y.)

Article Title: CXCL5 promotes prostate cancer progression.

doi: 10.1593/neo.07976

Figure Lengend Snippet: Figure 7. CXCL5 stimulates a transcriptional response in both nontransformed and transformed prostate epithelial cells. Quantitative real-time PCR of RNA purified from N15C6 cells (left) or LNCaP cells (right) treated with subnanomolar CXCL5 as shown demonstrates rapid and robust transcription of the EGR1 gene significantly higher than levels obtained at time 0 (set at one-fold) (*P < .05). Data shown are averaged from three or more separate experiments per time point per concentration of CXCL5 examined.

Article Snippet: To assess the effects of exogenous CXCL5 on cellular proliferation, recombinant human CXCL5 (254-X; R&D Systems) was added at the desired concentration in SF media (alone for control) to each well.

Techniques: Transformation Assay, Real-time Polymerase Chain Reaction, Purification, Concentration Assay

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: Nature Communications

Article Title: IL-17A is increased in diabetic wounds and impairs keratinocyte function via histone demethylase JMJD3

doi: 10.1038/s41467-025-67456-3

Figure Lengend Snippet: a ChIP analysis of H3K27me3 or IgG at indicated promoters in keratinocytes with (white) or without (blue) IL-17A stimulation. Itga3: n = 5 (unstimulated), n = 4 (IL-17A-stimulated) technical replicates, p = 0.0034, Timp1 : n = 3 technical replicates, p = 0.0046, Ccl20 : n = 3 technical replicates, p = 0.0059, Cxcl1 : n = 3 technical replicates, p = 0.0020, Cxcl3 : n = 3 technical replicates, p = 0.0258, Cxcl5 : n = 3 technical replicates, p = 0.0174. n = 3 independent experiments. b qPCR analysis of keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (1 µM) (red). Itga3 : n = 4 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0017 (IL-17A vs. IL-17A and inhibitor), Timp1: n = 6 biological replicates, p < 0.0312 (DMSO vs. IL-17A), p = 0.0029 (IL-17A vs. IL-17A and inhibitor), Ccl20: n = 3 biological replicates, p = 0.0008 (DMSO vs. IL-17A), p = 0.0428 (IL-17A vs. IL-17A and inhibitor), Cxcl1 : n = 3 biological replicates, p = 0.0003 (DMSO vs. IL-17A), p = 0.0294 (IL-17A vs. IL-17A and inhibitor), Cxcl3 : n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0003 (IL-17A vs. IL-17A and inhibitor), Cxcl5 : n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0007 (IL-17A vs. IL-17A and inhibitor), n = 3 independent experiments. c Western blot of ITGA-3 expression in keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (red). Representative densitometry plot is shown. n = 3 independent experiments. d Protein quantification of lysates from keratinocytes treated with DMSO only (white), with IL-17A alone (blue), or with IL-17A and GSK-J4 (red). TIMP-1: n = 6 biological replicates, p = 0.0012 (DMSO vs. IL-17A), p = 0.0068 (IL-17A vs. IL-17A and inhibitor), CCL-20: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0199 (IL-17A vs. IL-17A and inhibitor), CXCL-1: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0158 (IL-17A vs. IL-17A and inhibitor), CXCL-3: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0167 (IL-17A vs. IL-17A and inhibitor), CXCL-5: n = 3 biological replicates, p < 0.0001 (DMSO vs. IL-17A), p = 0.0005 (IL-17A vs. IL-17A and inhibitor), n = 3 independent experiments. e qPCR analysis of keratinocytes treated with a non-targeting control (siNTC) (white) or si Jmjd3 (gray). Jmjd3 : n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0192, Itga3: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0019, Timp1: n = 3 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0205, Ccl20: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0015, Cxcl1: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0019, Cxcl3: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0011, Cxcl5: n = 6 (siNTC), n = 4 biological replicates (si Jmjd3 ), p = 0.0011, n = 3 independent experiments. f qPCR analysis of Jmjd3 fl/fl K14 cre+ (red) and Jmjd3 fl/fl K14 cre- (yellow) keratinocytes. n = 3 biological replicates, Itga3 : p = 0.0184, Timp1 : p = 0.0044, Ccl20 : p = 0.0160, Cxcl1 : p = 0.0371, Cxcl3 : p = 0.0042. n = 3 independent experiments. g , h Scratch assays of primary murine ( n = 3 biological replicates, p < 0.0001 (48 h)) and N/TERT ( n = 3 biological replicates, p = 0.0340 (12 h)) keratinocytes treated with IL-17A alone (blue) or IL-17A and GSK-J4 (red). n = 3 independent experiments. i , j Scratch assays of primary murine ( n = 3 biological replicates, p = 0.0016 (48 h)) and N/TERT ( n = 6 biological replicates (IL-17A alone), n = 4 biological replicates (IL-17A and GSK-J1), p = 0.0223 (8 h), p < 0.0001 (12 h)) keratinocytes treated with IL-17A alone (blue) or IL-17A and GSK-J1 (red). n = 3 independent experiments. k Scratch assay of Jmjd3 fl/fl K14 cre+ (red) and Jmjd3 fl/fl K14 cre- (blue) keratinocytes. n = 3 biological replicates, p = 0.0198 (48 h). n = 3 independent experiments. Data were analyzed for variances, and 2-tailed Student’s t tests for ( a ), ( e ), ( f ) and 1-way ANOVA tests for ( b ), ( d ), ( g – k ) were performed. Data are presented as the mean ± SEM.

Article Snippet: After stimulation, cell free supernatant was collected and analyzed by the University of Michigan Immune Monitoring Shared Resource Core for CCL-20, CXCL-1, CXCL-5 or specific enzyme immunoassay kits for CXCL-3 (Boster Bio) and TIMP-1 (R&D Systems) according to the manufacturer’s instructions.

Techniques: Western Blot, Expressing, Control, Wound Healing Assay

Journal: bioRxiv

Article Title: Immune-Epithelial Interactions via TGF-β Orchestrates Stem-Cell Niche Formation and Morphogenesis

doi: 10.1101/2025.05.22.655596

Figure Lengend Snippet: (a) Cytokine array profiling comparing co-cultures and epithelial-alone hydrogels shows elevated levels of IL-6, CXCL5, CCL2, TIMP-1, LIF, GRO-a (CXCL1), IL-8, IGFBP-1, IGFBP-2, VEGF-A, SDF-1a, and IGF-1in responder co-cultures with immune cells (n=2 patients). (b) Bar graph showing treatment of responder epithelial cells with individual (IL-6, CCL2, GRO-a (CXCL1), CXCL5, VEGF) or pooled recombinant cytokines at 2ng/ml. (n=8; ns, not significant, **p-value < 0.01, ***p-value < 0.001). (c) Bar graph showing treatment of responder epithelium with conditioned media from responder co-cultures (n=8, ns, not significant). (d) Flow cytometry analysis of patient single cells showing percent of CD45 immune cells out of all breast cells. (n=12; ns, not significant). (e) Bar graph and scatter plot showing percent of CD3 + CD4 - CD8 - double negative (DN) T cells out of all CD45 + immune cells (n=12; ***p-value < 0.001, two-tailed t-test). (f) Scatter plot showing percent of γδ T cells out of all DN T cells.

Article Snippet: For treatment with recombinant proteins IL-6 (PHC0066, Thermofisher,2ng/ml), CCL2 (279-MC-050/CF, R&D Systems, 2ng/ml), GRO-a (275-GR-010/CF, R&D Systems, 2ng/ml), CXCL5 (254-XB-025/CF, R&D Systems, 2ng/ml), VEGF (78073, Stem Cell Technologies, 2ng/ml), treatment began at timepoint 0 and continued for the entire culture period.

Techniques: Recombinant, Flow Cytometry, Two Tailed Test

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: CXCL5 expression in the normal developing rat brain. a CXCL5 immunohistochemical analysis demonstrated that P7 ( n = 4) and, particularly, P2 pups ( n = 5) had significantly higher CXCL5 expression in the cerebral cortex than that of P30 rats ( n = 4). Values are means ± SEMs. The P2 pups had significantly higher CXCL5 expression in the cerebral cortex than that of the P7 pups and P30 rats. CXCL5 expression was low in the white matter, and no significant difference was observed between the three groups. b Immunofluorescence staining in the P2 pups revealed that the most CXCL5-positive cells in the cortex were neurons (NeuN) rather than microglia (ED1), and CXCL5 expression was absent in all white matter cells. c Scale bar = 50 μm

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Expressing, Immunohistochemical staining, Immunofluorescence, Staining

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: CXCL5 upregulation in the white matter after LPS-sensitized HI. a Western blots of the white matter revealed significant increases in CXCL5 levels at 6 and 24 h after LPS + HI ( n = 4 in each study group). b Immunohistochemical analyses revealed that the LPS + HI group had significantly increased CXCL5 immunoreactivity in the white matter at 6 h and, particularly, 24 h after HI compared with the control group ( n = 6 in each study group). Scale bar = 50 μm. Values are means ± SEMs. c Immunofluorescence analysis of the white matter 24 h after LPS + HI revealed that CXCL5 was upregulated mainly in activated microglia (ED1) and vascular endothelial cells (RECA) and not in astrocytes (GFAP) or preoligodendrocytes (O4). Scale bar = 50 μm. * p < 0.05, ** p < 0.01

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Western Blot, Immunohistochemical staining, Control, Immunofluorescence

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: Pharmacological inhibition of CXCR2 significantly attenuated microglial activation and BBB damage and protected against white matter injury after LPS-sensitized HI. A selective nonpeptide inhibitor of CXCR2, SB22 5002, was used to examine the role of the CXCL5–CXCR2 pathway in white matter injury after LPS-sensitized HI. After LPS + HI on P2, the SB-3 group ( n = 9), but not the SB-1 group ( n = 7), had significantly reduced ipsilateral ventricular size ratios ( a ), increased myelination (MBP) ( b ), and reduced astrogliosis (GFAP) ( c ) in the white matter on P12 compared with the vehicle-treated pups ( n = 9). Scale bar = 100 μm. The SB-3 group ( n = 9), but not the SB-1 group ( n = 7), had a significantly reduced number of activated microglia (ED1) ( d ) and significantly lower BBB damage (IgG extravasation) ( e ) in the white matter compared with the vehicle group ( n = 9) at 24 h after HI. Scale bar = 50 μm (ED1) and 100 μm (IgG); values are means ± SEMs, ** p < 0.01

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Inhibition, Activation Assay

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: CXCL5-sensitized HI induced microglial activation, neutrophil infiltration, and BBB damage and caused white matter injury. Intracerebroventricular infusion of recombinant CXCL5 (1 or 2 μg) or NS followed by HI on P2 showed that the 2-μg CXCL5 + HI group ( n = 6), but not the 1-μg CXCL5 + HI group ( n = 6), had significantly higher ipsilateral ventricle size ratios ( a ), lower myelination (MBP) ( b ), and higher astrogliosis (GFAP) ( c ) in the white matter compared with the NS ( n = 6) and the control groups ( n = 6) on P12. Scale bar = 100 μm. At 24 h after HI, the 2-μg CXCL5 + HI ( n = 6) and LPS + HI groups ( n = 6) had a significantly higher number of activated microglia (ED1) ( d ) and BBB damage ( e ) in the white matter compared with the NS ( n = 6) and control groups ( n = 6). The 2-μg CXCL5 + HI group, but not the LPS + HI group, had significantly increased neutrophil infiltrations (MPO) ( d ) in the white matter. Scale bar = 50 μm (ED1) and 100 μm (IgG and MPO); values are means ± SEMs, ** p < 0.01

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Activation Assay, Recombinant, Control

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: CXCL5 alone increased neutrophil infiltration and BBB damage and caused white matter injury. Intracerebroventricular infusion of recombinant CXCL5 (2 μg) or NS on P2 revealed that the CXCL5 group ( n = 6) had significantly higher ipsilateral ventricle size ratios ( a ), reduced myelination (MBP) ( b ), and increased astrogliosis (GFAP) ( c ) in the white matter on P12 compared with the vehicle group ( n = 6). Scale bar = 100 μm. ( d ) At 24 h after injection, the NS ( n = 6) and CXCL5 groups ( n = 6) showed no detectable ED1(+) activated microglia in the white matter. By contrast, the CXCL5 group had a significantly higher number of MPO(+) neutrophils and significantly greater BBB damage compared with the vehicle group. Scale bar = 50 μm (ED1) and 100 μm (IgG and MPO); values are means ± SEMs, * p < 0.05, ** p < 0.01

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Recombinant, Injection

Journal: Journal of Neuroinflammation

Article Title: CXCL5 signaling is a shared pathway of neuroinflammation and blood–brain barrier injury contributing to white matter injury in the immature brain

doi: 10.1186/s12974-015-0474-6

Figure Lengend Snippet: Diagram showing that the proposed CXCL5-mediated signaling pathway, triggered by LPS-sensitized HI, CXCL5-sensitized HI, or CXCL5 alone, plays a crucial role in neuroinflammation and BBB disruption and subsequent white matter injury in the immature brain. Neuroinflammation and BBB damage are two mutually potentiating mechanisms leading to sustained neuroinflammation and BBB disruption in white matter injury of the developing brain. White matter injury can be induced by different neuroinflammatory mechanisms: predominant microglial activation by LPS-sensitized HI, microglial activation and neutrophil infiltration by CXCL5-sensitized HI, and predominant neutrophil infiltration by CXCL5 alone

Article Snippet: The P2 pups were anesthetized using 2.5 % halothane and intracerebroventricularly infused with recombinant rat CXCL5 (25 μg of CXCL5 dissolved in 50 μL of NS, R&D Systems) or NS into the right cerebral hemisphere by using a 30-gauge needle on a 10-μL Hamilton syringe.

Techniques: Disruption, Activation Assay