Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Mo were cultured with or without the stimuli listed on the x-axis (concentrations as in Figure 1). Cell culture supernatants were collected at indicated time points and analyzed for the presence of CXCL16 by enzyme-linked immunosorbent assay (ELISA). Data represent the mean ± SEM from 3 independent experiments using 3 different donors with each condition tested in triplicate.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Mo were cultured with or without the indicated stimuli. Summary data are shown for the percent of CXCL16+ADAM10+ cells as a function of cell stimulus. Data are from 3 independent experiments using 3 different donors and are presented as the mean ± SEM.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Cell Culture

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Static adhesion of Mϕ to CASMCs, which were cultured with 25 µg/ml oxLDL for 24 h. The adhesion assay was performed as described in Materials and Methods. A, Prior to the adhesion assay, Mo or Mϕ were blocked for 45 min at 37°C with the following agents: 5 µg/ml isotype control rat IgG2b or CX3CR1 mAb or 4 µg/ml rat IgG2a or CXCL16 mAb. B, Mo were nucleofected with 150 nM of the indicated CXCL16-specific (CXCL16) or negative control (-Cmed) sRNAi and then cultured for 24 h in the presence or absence of atherogenic lipids. Data in A are from 3 independent experiments using Mo from 3 different donors with each condition tested in duplicate. Results in B are from 4 independent experiments using 4 different donors, each condition tested in triplicate. Data are expressed as the mean ± SEM.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Cell Culture, Cell Adhesion Assay, Control, Negative Control

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Uptake of Dil-oxLDL was performed as detailed in Materials and Methods. Cell-associated fluorescence was expressed as nanograms of Dil-oxLDL per milligram of total cellular protein. A, Prior to Dil-oxLDL internalization assay, Mϕ were incubated for 45 min at 37°C with 4 µg/ml of isotype control or mAbs recognizing CD36, SR-A, CXCL16 or CD68. B, Mo were nucleofected with 150 nM of the indicated CXCL16-specific (CXCL16) or negative control (-Cmed) sRNAi and then cultured for 24 h with or without 50 µg/ml oxLDL, 10 µg/ml 9-HODE or 10 µg/ml 13-HODE. Values shown are the mean (±SEM) of n=4 donors for each condition.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Fluorescence, Incubation, Control, Negative Control, Cell Culture

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Uptake of Dil-HDL was performed as described in Materials and Methods. Cell-associated fluorescence was expressed as nanograms of Dil-HDL per milligram of total cellular protein. A, Mϕ, which were differentiated in presence of 50 µg/ml oxLDL for 24 h, were incubated with 4 µg/ml rat IgG2a or CXCL16 mAb before Dil-HDL internalization assay. B, Mo were nucleofected with 150 nM of CXCL16-specific (CXCL16) or negative control (-Cmed) sRNAi and then cultured for 24 h with or without 50 µg/ml oxLDL, 10 µg/ml 9-HODE or 10 µg/ml 13-HODE. Data represent the mean ± SEM from 4 donors. *p < 0.05 and **p < 0.01 vs the corresponding unblocked control value in A.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Fluorescence, Incubation, Negative Control, Cell Culture, Control

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Mo were nucleofected with 150 nM of CXCL16-specific (CXCL16) or negative control (-Cmed) sRNAi and then cultured for 24 h with 50 µg/ml acLDL in the presence of radioactive cholesterol. Cells were equilibrated, washed and efflux was stimulated by incubating Mϕ in media containing lipid-poor acceptors for 6, 12 or 24 h. A, Macrophage 3H-Cholesterol efflux to human HDL. B, The release of 3H-Cholesterol to purified human ApoA-1. Results represent the mean ± SEM and are from 3 independent experiments using 3 different donors with each condition tested in triplicate. *p < 0.05, comparing the indicated value to the corresponding negative control sRNAi (-Cmed) value for the same time point.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Negative Control, Cell Culture, Purification

Journal:

Article Title: Atherogenic Lipids Induce HDL Uptake and Cholesterol Efflux in Human Macrophages by Upregulating Transmembrane Chemokine CXCL16, without Engaging CXCL16-dependent Cell Adhesion

doi: 10.4049/jimmunol.0804112

Figure Lengend Snippet: Mo were nucleofected with 150 nM of CXCL16-specific (CXCL16) or negative control (-Cmed) sRNAi and then cultured for 12, 24 or 48 h with or without 50 µg/ml oxLDL. Cells were harvested at indicated times, RNA isolated as described in Materials and Methods, and accumulation of mRNA for ABCA1 (A), ABCG1(B) and ApoE(C) was examined. Results represent the mean ± SEM and are from 5 independent experiments with each condition tested in triplicate. Donors were the same in A–C. *p < 0.05 and **p < 0.01 vs the corresponding negative control sRNAi (-Cmed) value for the same time point.

Article Snippet: Monoclonal antibodies (mAbs) included: rat anti-human CX3CR1 (Caltag-MedSystems Ltd, Buckingham, UK), rat anti-human CXCL16 (R&D Systems Europe Ltd, Abingdon, UK), mouse anti-human CD14, CD36, CD68 (BD Biosciences, Oxford, UK) and SR-A (CosmoBio, Tokyo, Japan).

Techniques: Negative Control, Cell Culture, Isolation

Journal: Genes

Article Title: Differentially Expressed Genes of Natural Killer Cells Can Distinguish Rheumatoid Arthritis Patients from Healthy Controls

doi: 10.3390/genes11050492

Figure Lengend Snippet: ELISA Quantification of ( A ) CXCL16 and ( B ) IL-1β and ( C ) IFN-γ in the plasma of RA patients and healthy controls.

Article Snippet: The kits used were human CXCL16 (DY1164), IFN-γ (DY285) and IL-1β (DY201) duosets (R&D systems, Minneapolis, MN, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 1. The transcription and translation of chemokine CXCL16 in human first-trimester trophoblasts. The real-time quantitative PCR was used to analyze transcription of CXCL16 in trophoblasts and JAR cells with cyclophilin A (CyP) as positive control. Three independent experiments were done (including 15 placental samples) and the results were reproducible (A). The specific brown-colored stainings for CXCL16 were recognized in the cytoplasm and cytomembrane of the primary-cultured villous cytotrophoblasts (B), and extravillous cytotrophoblasts (C) by immunocytochemistry, and the positive stainings were recognized by villous cytotrophoblasts and ST (E) and by extravillous cytotrophoblasts (F) by immunohistochemistry. No background staining was observed in goat isotype controls (D and G). The experiments were repeated five times with five placenta samples, respectively. The picture is a representative one. Magnification: B, D, E, and G, 200; C and F, 400.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Real-time Polymerase Chain Reaction, Positive Control, Cell Culture, Immunocytochemistry, Immunohistochemistry, Staining

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 2. The accumulated concentration of CXCL16 in culture me- dium of first-trimester cytotrophoblasts was examined by ELISA. Purified cytotrophoblasts were seeded at 1 106, 5 105, 1 105, 5 104, 1 104, and 5 103 cells/ml, and then the supernatants were collected and measured after 12, 24, 36, 48, 60, 72, and 100 h of culture. The results showed CXCL16 accumulation in the medium during the course of culture. Each point represents the mean SD obtained from duplicate dishes, and the data are representative of two independent experiments.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Human trophoblasts recruited T lymphocytes and monocytes into decidua by secretion of chemokine CXCL16 and interaction with CXCR6 in the first-trimester pregnancy.

doi: 10.4049/jimmunol.180.4.2367

Figure Lengend Snippet: FIGURE 6. Correlation of CXCL16 chemotaxis to CXCR6 expression level in the peripheral (A) and decidual (B) immune cells. CXCL16 che- motaxis was shown as the ratio of the number of migrated immune cells treated by CXCL16 (100 ng/ml) to the number of migrated immune cells in the control.

Article Snippet: The human CXCL16 ELISA kit (R&D Systems) was used to measure chemokine production in each supernatant and CM according to the manufacturer’s instructions.

Techniques: Chemotaxis Assay, Expressing, Control

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 1 CXCL1 is highly expressed in tissues with UCC. Strong immunohistochemical staining of CXCL1 in tissues with stage I (A), II (B) and III (C), comparing non-cancerous tissues with weak staining (D), was detected in a uterine cervical tissue microarray. Arrows, CXCL1 expression in stroma

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Immunohistochemical staining, Staining, Microarray, Expressing

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 2 High expression of CXCL1 positively correlated with worse survival in patients with CESC. A The role of CXCL1 expression level on patient overall survival probability. B The role of CXCL1 expression level and race on patient overall survival probability. C The role of CXCL1 expression level and body weight on patient overall survival probability. D The role of CXCR2 expression level on patient overall survival probability. E The role of CXCR2 expression level and race on patient overall survival probability. F The role of CXCR2 expression level and body weight on patient overall survival probability

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 3 CXCL1 expression was positively related with cancer-associated chemokines expression in CESC. A The genes that positively correlated with CXCL1 expression. B Heatmap of expression associated assay of 25 most relevant genes. C Gene expression correlation assays between CXCL and CCL20 (C), CXCL8 (D) or CXCL3 (E)

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing, Gene Expression

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 4 Exogenous CXCL1 facilitates the malignant behaviors of HeLa cells. A The role of different concentrations of exogenous CXCL1 on the proliferation of HeLa cells was tested by CCK-8 assay. B-C Cell migration ability was determined by transwell analysis after HeLa cells treatment with different concentrations of exogenous CXCL1. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: CCK-8 Assay, Migration

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 5 CXCL1 overexpression contributes to proliferation, migration and apoptosis of HeLa cells via a autocrine manner. A The protein expression of CXCL1 in the supernatant derived from HeLa cells overexpressing CXCL1 and their mock controls medium was assessed by ELISA assay. B The proliferation ability of HeLa cells overexpressing CXCL1 was significantly increased as compared to their mock controls. C-D The cell migration capacity was notably enhanced after CXCL1 overexpression in HeLa cells. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Over Expression, Migration, Expressing, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 6 Stromal cell-derived CXCL1 enhances oncogenic potential of HeLa cells via a paracrine manner. A CXCL1 protein level in PHM1-41 cells overexpressing CXCL1 and their mock controls-derived supernatant was determined using ELISA assay. B The growth ability of HeLa cells treatment with CM from PHM1-41 cells overexpressing CXCL1 and their mock controls was detected by CCK-8 assay. C-D The effect of various proportions of CM derived from PHM1-41 cells overexpressing CXCL1 and their mock controls on the HeLa cell migration was explored by transwell analysis. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Migration

Journal: BMC cancer

Article Title: High expression level of CXCL1/GROα is linked to advanced stage and worse survival in uterine cervical cancer and facilitates tumor cell malignant processes.

doi: 10.1186/s12885-022-09749-0

Figure Lengend Snippet: Fig. 7 ERK signal involves in CXCL1-mediated malignant phenotypes in HeLa cells. A Expression of ERK, p-ERK, Cyclin D1 and Bax at protein levels from HeLa cells overexpressing CXCL1 and their mock controls were assessed by western blotting. B The protein level of the p-ERK was quantitated and normalized to total ERK from western blotting. C Protein expression of ERK, Cyclin D1 and BAX were statistically analyzed from western blotting, and their bands were normalized to β-actin. D The proliferation inhibition rate of ERK inhibitor PD98059 on HeLa cells overexpressing CXCL1 and their mock controls was determined by CCK-8 assay. E–F The effect of ERK inhibitor PD98059 on migration inhibition rate of HeLa cells overexpressing CXCL1 and their mock controls was assessed by transwell analysis. *P < 0.05, **P < 0.01

Article Snippet: The culture supernatant derived from cell medium was collected and the concentration of the CXCL1 secretory protein was determined using Human CXCL1 ELISA Kit (Boster Biological Technology, China) following the manufacturer’s instruction.

Techniques: Expressing, Western Blot, Inhibition, CCK-8 Assay, Migration