Journal: Journal of neuroinflammation

Article Title: Astrocytic CXCL5 hinders microglial phagocytosis of myelin debris and aggravates white matter injury in chronic cerebral ischemia.

doi: 10.1186/s12974-023-02780-3

Figure Lengend Snippet: Fig. 1 Chronic cerebral ischemia induced significant up-regulation of CXCL5. a Volcano plot of the results of CC tissue microarray analysis between the Sham and BCAS groups at 2 months after surgery (fold change > 2; p < 0.05). n = 3 per group. b Heatmap of fourteen genes related to immune response form CC tissue microarray analysis. n = 3 per group. c The mRNA level of fourteen genes was measured in the Sham and BCAS groups by qPCR. n = 4 per group. d The mRNA level of Cxcl5 in the CC at different time points after BCAS by qPCR. e The result of serum CXCL5 concentration in the Sham and BCAS groups by ELISA. n = 8 per group. f The result of CXCL5 concentration in the CC homogenate in the Sham and BCAS groups by ELISA. n = 7–8 per group. g Representative images of immunostaining against GFAP and CXCL5 in EC in the BCAS groups. (Bar = 50 μm). h The mRNA level of Cxcl5 in the astrocytes (AST) and the other cells (Others) from the CTX and CC in the Sham and BCAS groups by qPCR. n = 4 per group. i The result of CXCL5 concentration in the supernatant of primary astrocytes in the control, OGD/R 12 h and OGD/R 24 h groups by ELISA. n = 3 per group. All data were presented as the mean ± SEM. *p < 0.05, ***p < 0.001, ns means no significance. EC: external capsule. 2W, 2 weeks. 2 M, 2 months. Student’s t-test for c, e and f. One-way ANOVA with Tukey’s post-hoc for d, i, j

Article Snippet: Enzyme linked immunosorbent assay (ELISA) The concentration of CXCL5 in mouse serum, CC homogenate and cell supernatant was quantified by ELISA kit (CUSABIO, China) according to the manufacturer’s instructions.

Techniques: Microarray, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunostaining, Control

Journal: Journal of neuroinflammation

Article Title: Astrocytic CXCL5 hinders microglial phagocytosis of myelin debris and aggravates white matter injury in chronic cerebral ischemia.

doi: 10.1186/s12974-023-02780-3

Figure Lengend Snippet: Fig. 2 Astrocyte specific Cxcl5 deficiency alleviated cognitive decline in BCAS model. a The result of CXCL5 concentration in the supernatant of primary astrocytes in the WT and Cxcl5 cKO groups after OGD/R 12 h by ELISA. n = 4 per group. b, c Results of the OFT showing the time spent in the corner area (b) and center area (c) in the WT and Cxcl5 cKO groups at 2 months after surgery. n = 12 per group. d Results of the NOR showing the recognition index percentage. n = 12 per group. e, f Results of Y-maze tests showing representative heatmaps (e) and the spontaneous alternation percentage (f). n = 12 per group. g–k Results of the MWM test. Representative heatmaps (g), escape latency during the acquisition phase (Days 1–5) (h), the time in target quadrant (i), the crossing platform times (j) and the swimming speed (k) of the probe test (Day 6). n = 11–12 per group. All data were presented as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns means no significance. Two-way ANOVA with Bonferroni’s post hoc test for a and h. Student’s t-test for b–d, j. Mann–Whitney test for f, i, k

Article Snippet: Enzyme linked immunosorbent assay (ELISA) The concentration of CXCL5 in mouse serum, CC homogenate and cell supernatant was quantified by ELISA kit (CUSABIO, China) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The clinic application of mNGS and ENA-78 assays to identify intra-amniotic infection/inflammation

doi: 10.3389/fcimb.2025.1510671

Figure Lengend Snippet: (A) Among the 40 preterm patients, the incidence of IAI was significantly higher than MIAC. (B) The ENA-78 concentration in AF of MIAC and IAI were higher than that of non-IAI/I, **p <0.01, **** p <0.0001. (C) The AUC of ENA-78 is 0.900, and the sensitivity and specificity was 73.3% and 100% respectively. (D-F) The NETs-makers concentration of AF was highest in MIAC group, middle in IAI group, and lowest in non-IAI/I group, **** p <0.0001. Data are presented as the mean ± SEM. Comparisons between the different groups were performed using Student’s t-test. MIAC, microbial invasion of the amniotic cavity; IAI, intra-amniotic inflammation; non-IAI/I, non-intra-amniotic infection/inflammation; ENA-78, Epithelial Neutrophil Activating Peptide-78; AUC, Area Under Curve; NETs, Neutrophil extracellular traps; cf-DNA, cell free-DNA; MPO, Myeloperoxidase; NE, neutrophil elastase; SEM, standard error of the mean.

Article Snippet: Human neutrophils (3×10 4 cells/ml) were plated on 96-well plates coated with poly-L-lysine (Solarbio,Beijing,China) and incubated with PMA (MedChemExpress, New Jersey, USA) or ENA-78 (MedChemExpress, New Jersey, USA), or leave untreated for 3 hours at 37 ° C in a 5% CO 2 atmosphere.

Techniques: Concentration Assay, Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The clinic application of mNGS and ENA-78 assays to identify intra-amniotic infection/inflammation

doi: 10.3389/fcimb.2025.1510671

Figure Lengend Snippet: ENA-78 promotes NETs generation. (A) Typical NETs structures were co-localized with MPO (red), NE (green), and DNA(blue). NETs production after ENA-78, PMA stimulation were significantly higher than NC, original magnification x 20, scale bars = 100μm. (B) Live-cell imaging shows NETs skeleton: cf-DNA, Sytox Green staining represents extracellular DNA; Hoechst 33342 staining represents intracellular DNA, original magnification x 10, Scale bars =100μm, ** P < 0.01; Data are presented as the mean ± SEM. Comparisons between the different groups were performed using Student’s t-test or one-way ANOVA. ENA-78, Epithelial Neutrophil Activating Peptide-78; NETs, Neutrophil extracellular traps; cf-DNA, cell free-DNA; MPO, Myeloperoxidase; NE, neutrophil elastase; DAPI:4’,6-diamidino-2-phenylindole; PMA, phorbol-12-myristate-13-acetate; NC, negative control; SEM, standard error of the mean.

Article Snippet: Human neutrophils (3×10 4 cells/ml) were plated on 96-well plates coated with poly-L-lysine (Solarbio,Beijing,China) and incubated with PMA (MedChemExpress, New Jersey, USA) or ENA-78 (MedChemExpress, New Jersey, USA), or leave untreated for 3 hours at 37 ° C in a 5% CO 2 atmosphere.

Techniques: Live Cell Imaging, Staining, Negative Control

Journal: PLoS ONE

Article Title: Necroptosis in pancreatic cancer promotes cancer cell migration and invasion by release of CXCL5

doi: 10.1371/journal.pone.0228015

Figure Lengend Snippet: (a) Images of human cytokine antibody array (120 targets) of conditioned medium derived from AsPC-1 treated with necroptotic stimuli or DMSO (control). Boxes: positive controls; circles: CXCL5, MIP-3α and IL-8. (b) Signals were quantified relative to CM-control. (c) Human cytokine antibody array of conditioned medium from PANC-1, which was treated with apoptotic stimuli or DMSO (control). (d) Signals relative to CM-control. (e, f) Analysis of mRNA expression of chemokine receptors, CXCR2 and CCR6 by qRT-PCR. Results are shown relative to gene expression in non-cancerous HPDE cells after normalization against 18S rRNA. (g) Western blot analysis of CXCR2 in human pancreatic cancer cells and in HPDE. (h) Concentration of CXCL5 in conditioned medium from AsPC-1 or BxPC-3, which were treated with TSZ ± nec-1 or DMSO (control), and measured by ELISA. Graphs show mean ± SE. * P < 0.05; ** P <0.01.

Article Snippet: After 12 hours’ incubation, CM was collected, and CXCL5 was quantified using a Human CXCL5 Quantikine ELISA Kit (R & D Systems Europe Ltd, Oxford, UK).

Techniques: Ab Array, Derivative Assay, Control, Expressing, Quantitative RT-PCR, Gene Expression, Western Blot, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: PLoS ONE

Article Title: Necroptosis in pancreatic cancer promotes cancer cell migration and invasion by release of CXCL5

doi: 10.1371/journal.pone.0228015

Figure Lengend Snippet: PC cell motility was evaluated by Transwell migration and Transwell-Matrigel invasion assays. PC cells pretreated with rh CXCL5 (10 ng/ml) or vehicle for 12 hours. (a) Representative images of migrated cells. scale bars = 100 μm. (b) Quantitative data of migrated cells. (c) Quantitative data of invaded cells. Graphs show mean ± SE. * P < 0.05; ** P <0.01.

Article Snippet: After 12 hours’ incubation, CM was collected, and CXCL5 was quantified using a Human CXCL5 Quantikine ELISA Kit (R & D Systems Europe Ltd, Oxford, UK).

Techniques: Migration

Journal: Advanced Science

Article Title: Tumor Microenvironment Responsive CD8 + T Cells and Myeloid‐Derived Suppressor Cells to Trigger CD73 Inhibitor AB680‐Based Synergistic Therapy for Pancreatic Cancer

doi: 10.1002/advs.202302498

Figure Lengend Snippet: AB680 regulates CXCL5 released by tumor cells and promotes MDSC accumulation. A) Heatmap of immune‐related differential genes in the RNA‐seq data of control and AB680‐treated tumors ( n = 3 samples per group). B,C) Over‐Representation Analysis B) and Gene Set Enrichment Analysis (C) of the above RNA‐seq data. D) Heatmap of 23 different mouse cytokine expression levels in serums obtained at the time of tumor collection, as shown in Figure using Bio‐Plex Pro Mouse Cytokine 23‐plex immunoassay ( n = 4 samples per group). E) The protein level of CXCL5 in mouse serums from the indicated treatment groups. Data presented as mean ± SD, ** p < 0.01 in Student's t ‐test. F) Dot plot of Cxcl5 expression in the identified populations in the single‐cell RNA‐sequencing dataset GSE158356 of two orthotopic KPC allografts. G,H) Quantification of CXCL5 and CXCR2% positive area in the AB680‐treated versus Control KPC tumor at the endpoint ( n = 6 samples per group). ** p < 0.01, *** p < 0.001 in the Student's t ‐test. Data presented as mean ± SD. I) Representative images of immunohistochemistry staining of CXCL5 and CXCR2 in the KPC tumors from the indicated treatment groups. J,K) The level of adenosine and CXCL5 protein in the AB680‐treated versus Control KPC tumor at the endpoint ( n = 6 samples per group). ** p < 0.01, *** p < 0.001 in Student's t ‐test. Data presented as mean ± SD. L) Schematic representation of the changes in adenosine metabolism after inhibition of CD73 activity by AB680. M) qRT‐PCR analysis of Cxcl5 mRNA expression in the KPC cells with different concentrations of AMP, adenosine, and the adenosine receptor agonist NECA at 0, 10, 20, 50, and 100 µ m ( n = 3). Data presented as mean ± SD. N) The CXCL5 protein concentration in the culture supernatant was evaluated by ELISA in the KPC cells with indicated concentrations of drugs ( n = 3). Data presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 in Student's t ‐test.

Article Snippet: Furthermore, the protein levels of CXCL5 and adenosine in KPC tumors and cell culture medium were detected using a mouse CXCL5 ELISA Kit (Boster, EK0919) and Adenosine Assay (Cell Biolabs, MET‐5090).

Techniques: RNA Sequencing, Control, Expressing, Immunohistochemistry, Staining, Inhibition, Activity Assay, Quantitative RT-PCR, Protein Concentration, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Tumor Microenvironment Responsive CD8 + T Cells and Myeloid‐Derived Suppressor Cells to Trigger CD73 Inhibitor AB680‐Based Synergistic Therapy for Pancreatic Cancer

doi: 10.1002/advs.202302498

Figure Lengend Snippet: CXCR2 inhibition further potentiates the efficacy of AB680 plus anti‐PD‐1 therapy. A) Schematic representation of the therapy schedule for AB680, anti‐PD‐1 antibody, and CXCR2 inhibitor SB225002 in the KPC orthotopic model. B,C) The image (B) and tumor weight (C) of KPC orthotopic allografts from the Control, SB225002, anti‐PD‐1+AB680, and combination groups ( n = 6/group). ** p < 0.01, *** p < 0.001 in Student's t ‐test. Data presented as mean ± SD. D) Survival of immunocompetent mice harboring KPC orthotopic allografts treated with the indicated treatment compared with the survival of those untreated mice ( n = 8/group). **p < 0.01, *** p < 0.001 in the log‐rank test. E) The percentages of total CD8 + T cells, Granzyme B+ cells among the total CD8 + T cells, and CXCR2 + MDSCs in the tumors of each group at the endpoint were determined by flow cytometry. n = 4 samples per group. Data presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 in Student's t ‐test. F) Quantification of CD8, Granzyme B, CXCR2, and Cleaved caspase‐3% positive area in the tumors of each group at the endpoint. n = 6 samples per group. Data presented as mean ± SD. ** p < 0.01, *** p < 0.001 in Student's t ‐test (G) schematic diagram depicting the mechanism by which CXCR2 inhibitor further potentiates the efficacy of AB680 plus anti‐PD‐1 therapy. AB680 can significantly increase the infiltration of responsive CD8+ T cells by suppressing the production of immunosuppressive adenosine, but it also promotes the MDSC chemotaxis by tumor‐derived CXCL5 in an AMP manner. Targeting the CXCL5/CXCR2 pathway with the CXCR2 inhibitor SB‐225002 can disrupt the “side effect” of AB680 and potentiate the efficacy of AB680 plus anti‐PD‐1 therapy. Data presented as mean ± SD, Student's t ‐test was used unless otherwise indicated.

Article Snippet: Furthermore, the protein levels of CXCL5 and adenosine in KPC tumors and cell culture medium were detected using a mouse CXCL5 ELISA Kit (Boster, EK0919) and Adenosine Assay (Cell Biolabs, MET‐5090).

Techniques: Inhibition, Control, Flow Cytometry, Chemotaxis Assay, Derivative Assay

Journal:

Article Title: A CXCL5- and bFGF-Dependent Effect of PDGF-B-Activated Fibroblasts in Promoting Trafficking and Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.1016/j.yexcr.2008.04.007

Figure Lengend Snippet: Protein array analyses were conduced by testing the cell culture supernatants of FB-PDGF-B/Ad5 versus FB-LacZ/Ad5 using a protein array set of RayBio® Human Angiogenesis Antibody Array I kit. CXCL5 (ENA-78) and bFGF were elevated 8-fold and 5-fold in the PDGF-B-aFB group, respectively, compared to the control fibroblast group. Expression of IFN-γ, RANTES and thrombopoietin (ThBo) was slightly decreased. Experiments were repeated three times, independently. Means ± SD were analyzed by ANOVA or student t-test.

Article Snippet: Goat anti-human PDGF-B antibody, goat anti-human CXCL5 (ENA-78) antibody, rabbit anti-human bFGF, and isotype matched control antibodies were purchased from R&D Systems.

Techniques: Protein Array, Cell Culture, Ab Array, Control, Expressing

Journal:

Article Title: A CXCL5- and bFGF-Dependent Effect of PDGF-B-Activated Fibroblasts in Promoting Trafficking and Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.1016/j.yexcr.2008.04.007

Figure Lengend Snippet: (A) Inhibition of BM-MSC invasion/migration by blocking antibodies against PDGF-B, CXCL5 and bFGF. Effects of blocking antibodies against PDGF-B, CXCL5 or bFGF on BM-MSC migration/invasion in 3D gel. Average percent of GFP+-BM-MSCs per HPF in the presence of FB-LacZ/Ad5 versus FB-PDGF-B/Ad5. Invasion/migration of BM-MSCs into collagen gels was significantly suppressed (**P<0.05). When recombinant PDGF-B or blocking antibodies were used in the collagen, they were added to the medium at the same concentration. Concentration of blocking antibody was within its neutralization dose (ND50), as titrated by the manufacturer, R&D Systems. All experiments were performed in duplicate and repeated three times, independently. Means ± SD were analyzed by ANOVA or student t-test. (B) Representative sections under 20X magnification were shown (order from left to right is the same as the bar graph). (C) Effect of exogenous CXCL5 and/or bFGF on BM-MSC invasion/migration into 3D gels. Invaded cell number of GFP+-BM-MSCs per HPF in the 3D gels in which FBs were not activated by PDGF-B was shown. CXCL5 and/or bFGF significantly promoted invasion/migration of BM-MSCs into 3D collagen gels compared with non-stimulated (−) group. Data were normalized by comparison with hrPDGF-B groups which were set as “100”. All experiments were performed in duplicate and repeated three times, independently. Means ± SD were analyzed by ANOVA or student t-test.

Article Snippet: Goat anti-human PDGF-B antibody, goat anti-human CXCL5 (ENA-78) antibody, rabbit anti-human bFGF, and isotype matched control antibodies were purchased from R&D Systems.

Techniques: Inhibition, Migration, Blocking Assay, Recombinant, Concentration Assay, Neutralization, Comparison

Journal:

Article Title: A CXCL5- and bFGF-Dependent Effect of PDGF-B-Activated Fibroblasts in Promoting Trafficking and Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.1016/j.yexcr.2008.04.007

Figure Lengend Snippet: (A) The effects of blocking antibodies against PDGF-B, CXCL5 or bFGF on myofibroblast differentiation of BM-MSCs in 3D gel. Average percentage of α-SMA+-BM-MSCs per HPF in 3D assays with FB-LacZ/Ad5 versus FB-PDGF-B/Ad5, respectively, under treatment with neutralization antibodies against PDGF-B, CXCL5 or bFGF. See Fig 3A for the control. A partial, but significant inhibition of differentiation was observed. Concentration of blocking antibody was within its neutralization dose (ND50), as titrated by the manufacturer, R&D Systems. All experiments were performed in duplicate and repeated three times, independently. Means ± SD were analyzed by ANOVA or student t-test. (B) Representative sections under 20X magnification were shown (see Fig 3B for the control).

Article Snippet: Goat anti-human PDGF-B antibody, goat anti-human CXCL5 (ENA-78) antibody, rabbit anti-human bFGF, and isotype matched control antibodies were purchased from R&D Systems.

Techniques: Blocking Assay, Neutralization, Control, Inhibition, Concentration Assay

Journal: Particle and Fibre Toxicology

Article Title: Inflammogenic effect of polyacrylic acid in rat lung following intratracheal instillation

doi: 10.1186/s12989-022-00448-z

Figure Lengend Snippet: Analysis of cytokines in BALF and HO-1 in lung tissue following intratracheal instillation. A CINC-1/CXCL1 concentration in BALF. B CINC-2/CXCL3 concentration in BALF. C CXCL5 concentration in BALF. D HO-1 concentration in lung tissue. The expressions of CINC-1/CXCL1, CINC-2/CXCL3 and CXCL5 in BALF in the exposed groups were persistently higher compared to the control group in a dose dependent-manner during 1 month after the exposure. Data are presented as mean ± SE (* p < 0.05 and ** p < 0.01 indicate that the values are significantly higher than control group. † p < 0.05 and †† p < 0.01 indicate that the values are significantly lower than control group.)

Article Snippet: Immunostaining for CXCL5 was performed with rabbit anti-mouse CXCL5 polyclonal antibody (1:200 dilution, bs-2549R; Bioss Inc., Woburn, MA, USA), while using the lung tissue samples from the 1.0 mg CL-PAA-exposure group of one month after intratracheal instillation.

Techniques: Concentration Assay

Journal: Particle and Fibre Toxicology

Article Title: Inflammogenic effect of polyacrylic acid in rat lung following intratracheal instillation

doi: 10.1186/s12989-022-00448-z

Figure Lengend Snippet: Description of chemokine genes related to ‘inflammatory response’ among 58 genes upregulated ≧ eightfold

Article Snippet: Immunostaining for CXCL5 was performed with rabbit anti-mouse CXCL5 polyclonal antibody (1:200 dilution, bs-2549R; Bioss Inc., Woburn, MA, USA), while using the lung tissue samples from the 1.0 mg CL-PAA-exposure group of one month after intratracheal instillation.

Techniques:

Journal: Particle and Fibre Toxicology

Article Title: Inflammogenic effect of polyacrylic acid in rat lung following intratracheal instillation

doi: 10.1186/s12989-022-00448-z

Figure Lengend Snippet: Representative images of CXCL5 immunostaining in lung tissue at 1 month after exposure to CL-PAA. A control lung (HE staining), B 1.0 mg CL-PAA-exposed lung (HE staining), C control lung (CXCL5 immunostaining), D 1.0 mg CL-PAA-exposed lung (CXCL5 immunostaining). Positive cells of CXCL5 immunostaining on 1.0 mg CL-PAA-exposed lung were mainly macrophages (blue arrow heads) (internal scale bar = 250 μm for all)

Article Snippet: Immunostaining for CXCL5 was performed with rabbit anti-mouse CXCL5 polyclonal antibody (1:200 dilution, bs-2549R; Bioss Inc., Woburn, MA, USA), while using the lung tissue samples from the 1.0 mg CL-PAA-exposure group of one month after intratracheal instillation.

Techniques: Immunostaining, Staining

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Chemokine (C-X-C motif) ligand 5 (CXCL5) levels were elevated in the synovial fluid (SF) of patients with periprosthetic joint infection (PJI), especially in the Gram-positive bacterial component-associated PJI (GPC-PJI) group. a) The expression level of CXCL5 in SF from patients with aseptic loosening (AL) and PJI was measured using protein-array assays. b) ImageJ software was used to quantify signal intensity of CXCL5 expression (two patients in the AL group and three patients in the PJI group). c) CXCL5 concentrations in SF of AL, GPC-PJI, and Gram-negative bacterial PJI (GNB-PJI) patients were measured by performing enzyme-linked immunosorbent assays. The expression of CXCL5 in the SF of PJI patients was significantly increased (four patients in the AL group, 12 patients in the GPC group, and three patients in the GNB group). Data are presented as the mean (standard error of the mean (SEM)) and analyzed using independent-samples t -test. *p < 0.05.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Infection, Expressing, Protein Array, Software

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Effects of lipoteichoic acid (LTA) treatment on chemokine (C-X-C motif) ligand 5 (CXCL5) expression in preosteoblasts and osteoblasts. a) Time-course analysis of CXCL5 expression following LTA stimulation. The preosteoblast cell line MC3T3-E1 was cultured in osteogenic induction medium (OIM) for 14 days to induce differentiation into osteoblasts. Cells were treated with LTA (0.1 μg/ml) for 24, 48, and 72 hours, and CXCL5 expression was measured at each timepoint. b) Dose-response analysis of CXCL5 expression in preosteoblasts and osteoblasts following treatment with LTA at concentrations of 0, 0.001, 0.01, 0.1, 1, and 10 μg/ml. CXCL5 expression levels positively correlated with increasing LTA concentrations in both cell types. Data are presented as mean (standard error of the mean (SEM)) and were analyzed using one-way analysis of variance followed by Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001. PBS, phosphate-buffered saline.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Expressing, Cell Culture, Comparison, Saline

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Chemokine (C-X-C motif) ligand 5 (CXCL5) inhibits receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation. a), c), and e) Immunofluorescence staining confirmed that CXCL5 (1 or 50 ng/ml) inhibits RANKL-induced differentiation of RAW264.7 cells into cathepsin K (CTSK)-positive osteoclast-like cells. b), d), and f) CXCL5 decreased the differentiation of RAW264.7 cells into tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like cells. Purple indicates TRAP expression, and cyan indicates nuclei. g) and h) Western blot analysis showed that CXCL5 (1 ng/ml) inhibited RANKL-induced differentiation of RAW264.7 cells by suppressing the transcription factor nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), resulting in decreased CTSK expression at 72 hours of treatment. Data are presented as the mean (standard error of the mean (SEM)) and analyzed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison test. *p < 0.05, **p < 0.01, ***p < 0.001. DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Comparison

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Chemokine (C-X-C motif) ligand 5 (CXCL5) attenuates receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation by inhibiting tumour necrosis factor receptor-associated factor 6 (TRAF6) downstream signalling through phospholipase Cγ2 (PLCγ2) and c-Fos. a) Schematic illustration depicting the experimental model, in which RAW264.7 cells were induced to undergo osteoclast differentiation by RANKL stimulation, with or without CXCL5 co-treatment, to investigate the regulatory role of CXCL5 in osteoclastogenesis and associated signalling pathways. b) and c) Western blot analysis was conducted to examine the time- and concentration-dependent effects of CXCL5 on RANKL-induced signalling in RAW264.7 cells. CXCR2 expression was positively correlated with CXCL5 concentration, and phosphorylation of PLCγ2 was reduced after 120 minutes of CXCL5 co-treatment, suggesting inhibition of TRAF6 downstream signalling. d) and e) CXCL5 co-treatment did not affect PLCγ1 signalling, indicating isoform specificity in the regulatory effect on PLCγ family proteins. f) and g) Western blot analysis of c-Fos and calcineurin A protein expression following CXCL5 treatment in RANKL-stimulated RAW264.7 cells. CXCL5 reduced c-Fos expression at 30 minutes post-stimulation, while having no significant impact at later timepoints. Calcineurin A levels remained unchanged across all conditions. Data are presented as mean (standard error of the mean (SEM)) and analyzed using one-way analysis of variance followed by Tukey’s multiple comparison test. *p < 0.05.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Western Blot, Concentration Assay, Expressing, Phospho-proteomics, Inhibition, Comparison

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Chemokine (C-X-C motif) ligand 5 (CXCL5) does not significantly alter mitogen-activated protein kinase (MAPK), nuclear factor-kappa B (NF-κB), signal transducer and activator of transcription (STAT), or focal adhesion kinase (FAK) signalling during receptor activator of nuclear factor kappa-B ligand (RANKL)-induced osteoclast differentiation. a) and b) To investigate whether CXCL5 modulates early intracellular signalling in osteoclastogenesis, RAW264.7 cells were stimulated with RANKL (50 ng/ml) to induce osteoclast differentiation, with or without co-treatment with recombinant CXCL5. Western blot analysis was conducted to examine the phosphorylation status of MAPK pathway components (p38, JNK, and ERK) at various timepoints. No significant changes in the p-p38/p38, p-JNK/JNK, or p-ERK/ERK ratios were observed following CXCL5 co-treatment. c) and d) Similarly, NF-κB signalling activity was assessed by analyzing phospho-p65 levels over time. CXCL5 had no significant effect on RANKL-induced p65 phosphorylation. e) to g) The effects of CXCL5 on other osteoclast-related signalling molecules, including STAT5, STAT6, and FAK, were also evaluated in both time- and dose-dependent settings. No significant changes were detected in STAT5/6 phosphorylation or FAK expression levels. Data are presented as mean (standard error of the mean (SEM)) and were analyzed using one-way analysis of variance followed by Tukey’s multiple comparison test.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Recombinant, Western Blot, Phospho-proteomics, Activity Assay, Expressing, Comparison

Journal: Bone & Joint Research

Article Title: CXCL5 suppresses osteoclastogenesis and protects against lipoteichoic acid-induced bone loss by modulating PLCγ2 and c-Fos signalling in gram-positive periprosthetic joint infection

doi: 10.1302/2046-3758.153.BJR-2025-0290.R1

Figure Lengend Snippet: Chemokine (C-X-C motif) ligand 5 (CXCL5) antibody treatment aggravates lipoteichoic acid (LTA)-induced trabecular bone loss in a mouse model. a) Representative cross-sectional and horizontal views of knee joint sections from LTA-treated mice with or without CXCL5 antibody treatment. Mice received intra-articular injections of phosphate-buffered saline (PBS; vehicle control) or LTA (10 mg/kg) on day 0 and daily intraperitoneal injections of PBS or CXCL5 antibody (2 μg/mouse) from day 0 to day 6 for a total of seven injections and were killed on day 7. The results of micro-CT revealed that CXCL5 treatment enhanced the bone loss caused by LTA. b) Quantitative results of micro-CT analysis. The medial and lateral subchondral bone plate and epiphyseal trabecular bone were analyzed separately. Data are presented as mean (standard error of the mean (SEM)). Statistical analysis was performed using two-way analysis of variance followed by Tukey’s multiple comparisons test. *p < 0.05. BV, bone volume; BS, bone surface; PI.Th, plate thickness; Po, porosity; Tb.N, trabecular number; Tb.Sp, trabecular spacing; Tb.Th, trabecular thickness; TV, tissue volume.

Article Snippet: CXCL5 concentrations in SF were further measured in triplicate using enzyme-linked immunosorbent assay (ELISA) (DY254; R&D Systems).

Techniques: Saline, Control, Micro-CT

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: 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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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: The TMA was then incubated overnight with primary CXCL5 antibody (Human CXCL5/ENA-78 MAb clone 33160; R&D Systems, Minneapolis, MN) at a dilution of 1:25.

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