recombinant human cxcl5 protein  (R&D Systems)

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: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 3. CXCL5 expression correlates positively with bone metastasis progression. (A) Serum samples were isolated from tumor-bearing mice [14 d after intracardiac (I.C.) injection, 3 × 104 cells/mouse], and protein-based arrays were analyzed as described by R&D Biosystems. Heatmap is presented. Cropped blot for CXCL5 is presented in the bottom panel. (B) CXCL5 level in serum from mice under different experimental conditions (sham/basal level vs. 14-d post-I.C. or postorthotopic implantation (1 × 104 cells/mouse). A mouse-specific CXCL5 enzyme-linked immunosorbent assay (ELISA) kit (R&D Biosystems) was used for determining CXCL5 levels. (C) Bone marrow (BM) cells were isolated from WT (BMWT) and mda-9−/− (BMKO) mice and stimulated with tumor cell–derived conditioned media for 12 h. Total cellular RNA was extracted, and qPCR was performed to detect mouse CXCL5 mRNA. Data are presented as fold-change relative to the unstimulated wild- type (BMWT) group. Different letters in two variables are statistically significant (P < 0.05). (D) CXCL5 levels in serum from tumor-bearing mice receiving “sham” (represented as “−“) or “WT” (represented as “+”) bone marrow (experiments described in Fig. 1E). *P < 0.01. (E) A total of 3 × 104 RM1-BM-Luc cells were injected by the intracardiac (I.C.) route into WT and mda-9−/− mice (schematically presented in the left panel). Fourteen days postinjection, total BM cells were isolated and stained for lineage-specific cell surface markers and intracellular CXCL5 expression to identify the specific cell population(s) that differentially respond to tumor cells. Percentages of CXCL5 expression in bone marrow–derived mesenchymal stromal cells: (CD45−CD90+) are presented. *P < 0.05. (F) Tumor-implanted bones (from animals in Fig. 2A) were immunostained with different antibodies as indicated, and representative photographs are presented.

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Expressing, Isolation, Injection, Enzyme-linked Immunosorbent Assay, Derivative Assay, Staining

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 4. MDA-9 expression in tumor cells nonautonomously activates the Hippo pathway and induces CXCL5 in HS5 cells. (A) Bone metastasis development in athymic nude mice following intracardiac injection of PC-3ML control and mda-9 KO (PC-3MLmda-9 KO) clone (1 × 105 cells/mouse). Representative images 36 d after tumor cell implantation (Left). Bone metastases detected by BLI imaging. The percentage of mice with BLI-positive lesions is shown (Right). (B) HS5 cells were incubated with normalized (equal amount of total protein) tumor cell–derived condition media for 12 h. HS5-derived conditioned media were analyzed for CXCL5 levels using ELISA. (C) HS5 cells were transfected with a 1.5-kb CXCL5-Prom using a standard transfection protocol. Twelve hours posttransfection, cells were stimulated with conditioned media for an additional 12 h. Luciferase activity was measured and presented after normalizing with Renilla luciferase. (D) HS5 cells were incubated with tumor cell–derived conditioned media for 12 h and stained with fluorescently labeled anti-YAP. DAPI was used for nuclear staining. Translocation of YAP into the nucleus was monitored under a fluorescence microscope. (E) HS5 cells were incubated with tumor cell– (parental and its mda-9 knockout variant) derived conditioned media for 12 h, and cell lysates were analyzed for different proteins, as indicated. (F) MDA-9/Syntenin expression was knocked down in HS5 cells using an Adenovirus expressing shmda-9. Twenty-four hours postinfection, culture media were replaced with fresh media containing PC-3ML CM for an additional 12 h. Cell lysates were analyzed for the indicated proteins. Different letters in two variables are statistically significant (P < 0.05).

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Expressing, Injection, Control, Imaging, Incubation, Derivative Assay, Enzyme-linked Immunosorbent Assay, Transfection, Luciferase, Activity Assay, Staining, Labeling, Translocation Assay, Fluorescence, Microscopy, Knock-Out, Variant Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 6. Tumor cell–derived PDGF-AA induces CXCL5 expression in HS5 cells. (A) Normalized (based on equal amounts of total protein) conditioned media, isolated from parental and knockout variants, were subjected to protein-based growth factor arrays as described by Ray Biotech. Heatmap is presented. Densitometry analyses (n = 2, for corresponding proteins) were performed for each growth factor, and the data are presented only for three. (B) HS5 cells were treated for the indicated times with PDGF-AA, and CXCL5 protein expression was determined. (C) HS5 cells were transfected with a 1.5-kb CXCL5-Prom using a standard transfection protocol. Twelve hours posttransfection, cells were treated with PDGF-AA for an additional 12 h. Luciferase activity was measured and presented after normalizing with Renilla luciferase. (D) PC-3ML-derived conditioned media were incubated with control IgG or anti-PDGF-AA before treatment of HS5 cells for 12 h, and CXCL5 levels were measured in media. (E) Expression of PDGF-AA was determined in the indicated tumor cells and corresponding mda-9 knockout clones. (F) Cells were infected with an Adenovirus expressing control or mda-9 overexpressing construct for 24 h, and PDGF-AA expression in media was determined using ELISA. (G) HS5 cells were treated with PDGF-AA and analyzed for expression of the indicated proteins. (H) Tumor cells were cultured in the presence or absence of IKK2i for 12 h and analyzed for expression of the indicated proteins. Different letters in two variables are statistically significant (P < 0.05). (I) MDA-9 was knocked-into PC-3MLmda-9 KO cells and cultured for an additional 12 h in the presence or absence of IKK2i. PDGF-AA was determined using ELISA. *=P < 0.05.

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Derivative Assay, Expressing, Isolation, Knock-Out, Transfection, Luciferase, Activity Assay, Incubation, Control, Clone Assay, Infection, Construct, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 5. Hippo pathway activates CXCL5 in HS5 cells. (A) Panel i Schematic representation of the experimental protocol used in panels ii–iv. Different overexpression/ siRNAs (GOF/LOF) constructs were transfected into the indicated cells, and after 24 h, tumor-derived condition medium was used to stimulate HS5 cells for an additional 12 h. Panels ii and iii represent CXCL5 expression in HS5-derived conditioned media, determined by ELISA. Panels iv and v show CXCL5 promoter activity. In this experiment, HS5 cells were cotransfected with both CXCL5-Prom and either shYAP1 or YAP1 OE vectors before treating with conditioned media. *=P <0.05. (B) YAP expression was analyzed in paraffin sections (tumor-bearing bones from the experiment performed in Fig. 2A). (C) HS5 cells were incubated with different fractions of PC-3ML-derived conditioned media for 24 h after initial transfection with a CXCL5-Prom. Luciferase activity was measured and presented after normalizing with Renilla luciferase. *P <0.05.

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Over Expression, Construct, Transfection, Derivative Assay, Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Incubation, Luciferase

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 7. MDA-9 expression in HS5 regulates PDGF/PDGFRα signaling. (A) mda-9 expression was blunted, and 24 h postinfection, HS5 cells were stimulated with PDGF-AA for an additional 12 h. mRNA for CXCL5 expression was analyzed using qPCR. Different letters in two variables are statistically significant (P < 0.05). (B) HS5 cells were incubated with media supplemented with the indicated tumor cell–derived conditioned media for 12 h, and western blotting was done for phosho-PDGFRα and total PDGFR. (C) HS5 cells were incubated with media supplemented with the indicated tumor cell–derived conditioned media and PDGF- AA for 12 h, and western blotting was done for the indicated proteins. (D) BM-MSCs from WT and mda-9−/− mice were stimulated with mouse PDGF-AA, and CXCL5 expression was determined in the culture media. (E) phospho-PDGFR expression was analyzed in paraffin sections (tumor-bearing bone from the animal experiment, described in Fig. 2A). Different letters in two variables are statistically significant (P < 0.05).

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Expressing, Incubation, Derivative Assay, Western Blot

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 8. CXCL5 is expressed in both stroma and tumor cells. (A) Paraffin-embedded section from a patient with metastatic PC bone metastasis was immunostained with anti-CXCL5, and the expression in the stroma and tumor compartment is presented in the Inset. (B) H-Score was determined by an anatomic pathologist based on staining intensity, and values are presented. (C) PC3-ML cells were stained with CytoTrack™ Red and cocultured with of HS5 cells. After 24 h, cells were stained with CXCL5-FITC and analyzed by FACS [the left panel represents double staining (tumor cell-Red and CXCL5-FITC)]; the middle panel is for only CXCL5- FITC (only HS5). The right panel represents conditioned media that were collected from the above indicated PC3-ML and HS5 cocultured cells, and secretion of CXCL5 was measured by ELISA. (D) Human serum samples from the indicated patient groups were analyzed for PDGF-AA and CXCL5 (n = 20). Different letters in two variables are statistically significant (P < 0.05). *Statistically significant. **P < 0.001.

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Expressing, Staining, Double Staining, Enzyme-linked Immunosorbent Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: MDA-9/Syntenin in the tumor and microenvironment defines prostate cancer bone metastasis.

doi: 10.1073/pnas.2307094120

Figure Lengend Snippet: Fig. 9. Schematic of the proposed role of MDA-9 in BM- MSCs in creating a favorable environment for metastatic outgrowth through secretion of CXCL5 in response to tumor cell–derived growth factor, PDGF-AA.

Article Snippet: Human and mouse specific recombinant protein CXCL5, PDGF- AA, GM- CSF were procured from R&D Systems.

Techniques: Derivative Assay

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a QRT-PCR analyses of CXCL5 expression in gastric cancer tissues and non-tumor tissues. b QRT-PCR analyses of CXCL5 expression in gastric cancer cell lines and normal gastric epithelial cell line. c ELISA assays for CXCL5 concentrations in culture supernatants from gastric cancer cell lines and normal gastric mucosa epithelial cell line. d ELISA assays for CXCL5 concentrations in the serum of gastric cancer patients and healthy controls. e ELISA assays for CXCL5 concentrations in culture supernatants from tumor tissues and non-tumor tissues.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: The association between CXCL5 expression levels and clinicopathological factors in gastric cancer patients.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Expressing

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a Transwell migration and Matrigel invasion assays for GC cells treated with or without CXCL5. b , c QRT-PCR ( b ) and western blot ( c ) analyses of the expression of EMT markers in GC cells treated with or without CXCL5. d Transwell migration and Matrigel invasion assays for GC cells treated with culture supernatants from tumor tissues (TTCM) and normal tissues (NTCM) in the presence or absence of CXCL5 neutralizing antibody. e Western blot assays for the expression of EMT markers in GC cells treated with TTCM in the presence or absence of CXCL5 neutralizing antibody.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Migration, Quantitative RT-PCR, Western Blot, Expressing

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a Western blot assays for the expression of phosphorylated ERK in GC cells treated with CXCL5 for different times. b Transwell migration and Matrigel invasion assays for GC cells treated with CXCL5 in the presence or absence of ERK inhibitor PD98059. c Western blot assays for the expression of EMT markers in GC cells treated with CXCL5 in the presence or absence of ERK inhibitor PD98059. d Western blot assays for the expression of phosphorylated ERKs in GC cells treated with TTCM in the presence or absence of CXCL5 neutralizing antibody. e Transwell migration and Matrigel invasion assays for GC cells treated with TTCM in the presence or absence of ERK inhibitor PD98059.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Western Blot, Expressing, Migration

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a Chemotaxis assay for neutrophils induced by different concentrations of CXCL5 (10 and 100 nM). b Western blot assays for the expression of phosphorylated ERK and P38 in neutrophils treated with CXCL5. c QRT-PCR analyses of the expression of IL-6, IL-8, IL-17, IL-23, TNF-α, TGF-β, MMP9, VEGF, and PD-L1 in neutrophils treated with CXCL5. d Western blot assays for the expression of phosphorylated ERK and P38 in neutrophils treated with TTCM in the presence of CXCL5 neutralizing antibody. e QRT-PCR analyses of the expression of IL-6, IL-8, IL-17, IL-23, TNF-α, TGF-β, MMP9, VEGF, and PD-L1 in neutrophils treated with TTCM in the presence of CXCL5 neutralizing antibody. f , g QRT-PCR analyses of the expression of IL-6, IL-8, IL-17, IL-23, TNF-α, TGF-β, MMP9, VEGF, and PD-L1 in CXCL5-treated neutrophils in the presence or absence of ERK inhibitor PD98059 ( f ) and p38 inhibitor SB203580 ( g ).

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Chemotaxis Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a Transwell migration and Matrigel invasion assays for GC cells treated with conditioned medium of untreated (0-NCM) and CXCL5 (100 nM)-treated (100-NCM) neutrophils. b Western blot assays for the expression of EMT markers in GC cells treated with conditioned medium of untreated and CXCL5-treated neutrophils. c Transwell migration and Matrigel invasion assays for GC cells treated with conditioned medium of untreated (NCM) or TTCM-primed neutrophils (TNCM) in the presence or absence of CXCL5 neutralizing antibody. d ELISA assays for IL-6 and IL-23 concentrations in culture supernatants from CXCL5-primed neutrophils. e Transwell migration and Matrigel invasion assays for GC cells treated with conditioned medium of CXCL5-activated neutrophils in the presence or absence of IL-6 and IL-23 neutralizing antibodies. f Western blot assays for the expression of EMT markers in GC cells treated with conditioned medium of CXCL5-activated neutrophils in the presence or absence of IL-6 and IL-23 neutralizing antibodies.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Migration, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: a The number of tumor colonized into the abdomen and the metastatic nodules in liver of mice in control, CXCL5, 0-NCM, and 100-NCM groups. b , c H&E staining and immunohistochemical staining of liver tissues for hepatic metastases and E-cadherin in mice from control, CXCL5, 0-NCM, and 100-NCM groups. c , d QRT-PCR ( c ) and western blot ( d ) analyses of the expression of EMT markers in mouse metastatic tumor nodules. Scale bar: 50 μm.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Control, Staining, Immunohistochemical staining, Quantitative RT-PCR, Western Blot, Expressing

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: High level of CXCL5 in tumor microenvironment promotes GC metastasis by enhancing GC cell migration and invasion through the induction of EMT. CXCL5 exerts these effects by two mechanisms: (1) CXCL5 directly activates ERK signaling pathway in GC cells to upregulate Snail expression; (2) CXCL5 induces pro-tumor activation of neutrophils via ERK and p38 signaling pathways, which leads to the release of increased levels of inflammatory factors (e.g, IL-6 and IL-23), subsequently enhancing the metastatic potential of gastric cancer cells.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Migration, Expressing, Activation Assay, Protein-Protein interactions

Journal: Oncogenesis

Article Title: CXCL5 promotes gastric cancer metastasis by inducing epithelial-mesenchymal transition and activating neutrophils

doi: 10.1038/s41389-020-00249-z

Figure Lengend Snippet: The sequences of primers for target genes.

Article Snippet: Recombinant human CXCL5 (R&D Systems) was used to stimulate gastric cancer cells (HGC: 20 ng/mL, BGC: 60 ng/mL, 24 h) and neutrophils (100 ng/mL, 12 h).

Techniques: Sequencing

Journal: The Journal of investigative dermatology

Article Title: CXCL5 Facilitates Melanoma Cell-Neutrophil Interaction and Lymph Node Metastasis.

doi: 10.1016/j.jid.2018.01.035

Figure Lengend Snippet: Figure 1. CXCL5 is strongest up-regulated in high risk human cutaneous melanoma. (a) Fold change (FC) expression in primary melanomas (T1, n ¼ 18; T2, n ¼ 21; T3, n ¼ 19; T4, n ¼ 21) and nevi (N, n ¼ 11), normalized to normal skin (n ¼ 8). P values shown for differences between T1 and T4 melanomas. (b) FC of CXCL5 mRNA levels of lymph node metastasis normalized to primary tumor. (c) CXCR2 mRNA expression (DCt) in T1 (n ¼ 7), T2 (n ¼ 7), T3 (n ¼ 7) and T4 (n ¼ 13) melanomas. (d) Left: Positive correlation of DCt CXCL5 to CXCR2 mRNA expression in T4 melanomas (n ¼ 13). Right: FC of CXCR2 in CXCL5high (n ¼ 6) compared to CXCL5low (n ¼ 7) T4 melanomas. (e) Representative immunohistochemical stainings of CXCR2 in healthy skin and primary melanoma. Insert: IgG2a isotype control. Scale bar ¼ 200 mm. Mean standard error of mean (c, d); one-way analysis of variance, Tukey post-hoc test or two-tailed t test (c, d). *P < 0.05.

Article Snippet: Spheroids were embedded into a collagen/methocel matrix and stimulated with recombinant human CXCL5 (#254-XB, R&D Minneapolis, MN) with or without mouse anti-human CXCR2 blocking antibody Clone 48311 (#MAB331, R&D Minneapolis, MN), supernatants from VEGFC over-expressing cells as positive control or IMDM/3%BSA as negative control.

Techniques: Expressing, Immunohistochemical staining, Control, Two Tailed Test

Journal: The Journal of investigative dermatology

Article Title: CXCL5 Facilitates Melanoma Cell-Neutrophil Interaction and Lymph Node Metastasis.

doi: 10.1016/j.jid.2018.01.035

Figure Lengend Snippet: Figure 2. CXCL5 expression increases lymph node metastasis in a mouse xenograft model. (a) Left: Proliferation assay of M24 and A375 in vitro. Right: Representative pictures of an in vitro scratch assay for A375. (b) Growth curves of M24 and A375 xenografts in severe combined immune-deficient mice (n ¼ 5 per group, two independent experiments). (c) Enzyme-linked immunosorbent assay of CXCL5 serum levels in mice, carrying either M24 control (n ¼ 2) or M24 CXCL5ox (n ¼ 2) primary tumors. (d) Frequencies of lymph node metastases in control and CXCL5ox mice. Representative staining of anti-human VIMENTIN-stained lymph node micrometastasis. Scale bar ¼ 100 mm. (e) Weight (in milligrams) of tumor draining lymph nodes. M24 control, n ¼ 14, M24 CXCL5ox, n ¼ 14, A375 control, n ¼ 12, A375 CXCL5ox, n ¼ 14, three independent experiments (d, e). Mean standard error of mean (a, b, c, e); statistical analysis: two-tailed t test (c, e), c2 test (d). *P < 0.05.

Article Snippet: Spheroids were embedded into a collagen/methocel matrix and stimulated with recombinant human CXCL5 (#254-XB, R&D Minneapolis, MN) with or without mouse anti-human CXCR2 blocking antibody Clone 48311 (#MAB331, R&D Minneapolis, MN), supernatants from VEGFC over-expressing cells as positive control or IMDM/3%BSA as negative control.

Techniques: Expressing, Proliferation Assay, In Vitro, Wound Healing Assay, Enzyme-linked Immunosorbent Assay, Control, Staining, Two Tailed Test

Journal: The Journal of investigative dermatology

Article Title: CXCL5 Facilitates Melanoma Cell-Neutrophil Interaction and Lymph Node Metastasis.

doi: 10.1016/j.jid.2018.01.035

Figure Lengend Snippet: Figure 4. Lymphangiogenic potential of CXCL5. (a) Percentages of LYVE-1 and CD31-positive stained areas in sections of primary CXCL5ox and control tumors (n ¼ 6, two independent experiments). Scale bar ¼ 100 mm. (b) FACS histogram of lymphatic endothelial cells stained against CXCR2. One representative out of three independent experiments. (c) Cumulative length (103 mm) of sprouts from immortalized human lymphatic endothelial cells spheroids stimulated with rCXCL5 anti-CXCR2, VEGFC, or bovine serum albumin. One representative out of three independent experiments. Representative photographs of sprouting spheroids stimulated with rCXCL5 or VEGFC. (d) Total numbers of A375 melanoma cells transmigrated across a fibronectin-coated lymphatic endothelial cell monolayer in the presence or absence of neutrophils. One representative out of two independent experiments. Statistical analysis by two-tailed t test (a, d); mean standard error of mean (a, d). *P < 0.05.

Article Snippet: Spheroids were embedded into a collagen/methocel matrix and stimulated with recombinant human CXCL5 (#254-XB, R&D Minneapolis, MN) with or without mouse anti-human CXCR2 blocking antibody Clone 48311 (#MAB331, R&D Minneapolis, MN), supernatants from VEGFC over-expressing cells as positive control or IMDM/3%BSA as negative control.

Techniques: Staining, Control, Two Tailed Test

Journal: The Journal of investigative dermatology

Article Title: CXCL5 Facilitates Melanoma Cell-Neutrophil Interaction and Lymph Node Metastasis.

doi: 10.1016/j.jid.2018.01.035

Figure Lengend Snippet: Figure 5. CXCL5-expressing human primary melanomas have a higher risk for metastasis. (a) CD66bþ neutrophils per mm2 in CXCL5-positive (n ¼ 27) or -negative (n ¼ 13) histological sections of human melanoma samples. Representative images of anti-CD66bestained neutrophils on human melanomas, classified as CXCL5high or CXCL5low. Scale bar ¼ 100 mm. (b) CD66bþ Neutrophils per mm2 in histological sections of human melanoma samples graded in ulcerated (n ¼ 27) or not ulcerated (n ¼ 36). (c) Log2 fold-change expression of CXCL5 (mRNA) in thick human primary melanomas (tumor stage T3/T4) of patients with (n ¼ 14) or without (n ¼ 7) metastases. Statistical analysis by two-tailed t test and data are represented as box plot with individual data points. *P < 0.05.

Article Snippet: Spheroids were embedded into a collagen/methocel matrix and stimulated with recombinant human CXCL5 (#254-XB, R&D Minneapolis, MN) with or without mouse anti-human CXCR2 blocking antibody Clone 48311 (#MAB331, R&D Minneapolis, MN), supernatants from VEGFC over-expressing cells as positive control or IMDM/3%BSA as negative control.

Techniques: Expressing, Two Tailed Test