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: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

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: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

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: Anti-human CXCL16 Ab (R&D Systems) were administrated to detect whether cytotrophoblasts express CXCL16.

Techniques: Chemotaxis Assay, Expressing, Control

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: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Baseline clinical and laboratory data and levels of urinary CXCL1 in patients with IgAN.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Urinary CXCL1 was calibrated against urine creatinine before the levels were compared. The urinary CXCL1 levels in patients with IgAN, MCD, MN, FSGS and LN were significantly higher than those in healthy controls (0, IQR 0–0, P<0.001 in all glomerulonephritis). Urinary CXCL1 levels were significantly higher in patients with IgAN (18.29 pg/mg, IQR 10.17–33.47 pg/mg) than in MCD (3.03 pg/mg, IQR 0–11.28 pg/mg, P<0.001), MN (5.89 pg/mg, IQR 0–23.54 pg/mg, P<0.001) and FSGS (6.10 pg/mg, IQR 0–17.53 pg/mg, P<0.001), while no significant difference was observed between patient with IgAN and LN (18.29 pg/mg, IQR 10.17–33.47 pg/mg vs. 23.12 pg/mg, IQR 7.41–48.17 pg/mg; P = 0.873). (*, significant difference between IgAN group and other GN groups; **, significant difference between healthy control group and disease groups).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Control

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Multivariable linear regression analyses for urinary CXCL1 level.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: For patients with high baseline urinary CXCL1 levels (case 1 & case 2), the variation tendencies of urinary CXCL1 and proteinuria were almost the same (A & B). For patients with middle levels of baseline urinary CXCL1 (case 3 & case 4), although proteinuria relieved a lot, urinary CXCL1 levels just decreased a little (C & D). For patients with low levels of baseline urinary CXCL1 (case 5 & case 6), decreased urinary CXCL1 levels were not observed accompanied with proteinuria remission (E & F). Treatments are depicted as grey-level-coded bars, on behalf of the period of using time.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: The variation tendency of urinary CXCL1 was not the same with that of SBP, not only in patients with high baseline urinary CXCL1 levels (case 1 & case 2, A & B), but also in patients with middle baseline urinary CXCL1 levels (case 3 & case 4, C & D) and patients with low baseline urinary CXCL1 levels (case 5 & case 6, E & F). Treatments are depicted as grey-level-coded bars, on behalf of the period of using time.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Clinical outcomes according to urinary CXCL1 level.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Patients with IgAN were divided into two groups, according to the median urinary CXCL1 level (18.29 pg/mg). Those with CXCL1 levels less than 18.29 pg/mg were classified as group A, while others (CXCL1 levels above 18.29 pg/mg) were classified as group B. IgAN patients in group B had significantly lower renal survival rate than those in group A (P < 0.001). The renal survival at first and fifth year for patients in group A were 100.0% and 96.3%, while for patients in group B were 97.2% and 81.3% (Log Rank test, p<0.001).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Risks of composite end-point of natural log–transformed CXCL1.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques:

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Using composite outcome at 24 (solid line), 48 (dashed line), and 72 months (dotted line) as status variable, respectively, the areas under the ROC curve (AUC) for proteinuria (A), reciprocal of estimated glomerular filtration rate (1/eGFR) (B) and urinary CXCL1 (C) were comparable (proteinuria: 0.720 (24 mo), 0.690 (48 mo), 0.612 (72 mo); 1/eGFR: 0.826 (24 mo), 0.809 (48 mo), and 0.690 (72 mo); urinary CXCL1: 0.770 (24 mo), 0.651 (48 mo), 0.668 (72 mo)). When combined urinary CXCL1 level with proteinuria and 1/eGFR (solid line), the AUCs were higher than proteinuria alone (dotted line) or proteinuria plus 1/eGFR (dashed line) (D).

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Filtration

Journal: PLoS ONE

Article Title: Urinary CXCL1: A Novel Predictor of IgA Nephropathy Progression

doi: 10.1371/journal.pone.0119033

Figure Lengend Snippet: Comparison of two models of survival analysis.

Article Snippet: Urinary CXCL1 were quantified by a standard sandwich ELISA assays using the DuoSet human CXCL1 ELISA kits (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s protocol.

Techniques: Comparison

Journal: The Journal of infectious diseases

Article Title: CXCL16 contributes to neutrophil recruitment to cerebrospinal fluid in pneumococcal meningitis.

doi: 10.1086/656532

Figure Lengend Snippet: Figure 1. In vitro and in vivo expression analysis of CXCL16. A, Levels of soluble CXCL16 (expressed as nanograms per milliliter) in cerebrospinal fluid samples of patients with pneumococcal meningitis (right) ( ) and control patients with noninflammatory central nervous system diseases n p 20 (left) ( diagnoses: migraine [ ], tension-type headache [ ], exertional headache [ ], normal pressure hydrocephalus [ ], facial n p 16 n p 8 n p 1 n p 1 n p 1 paresis [ ], amyotrophic lateral sclerosis [ ], and herniated disc [ ]. Data are the mean value standard deviation ( ). B, CXCL16 n p 4 n p 1 n p 1 P ! .05 levels (expressed as picograms per milliliter) in the supernatant of cell cultures (RAW264.7 macrophages, bEnd5 brain microvascular endothelial cells, and Hoxb8 neutrophils; in each experiment) stimulated with ethanol-killed Streptococcus pneumoniae (EKP) or left unstimulated (medium). Data n p 6 are the mean value standard deviation ( ). C–E, Immunohistochemical staining of CXCL16 in formalin-fixed and paraffin-embedded brains: P ! .05 uninfected control mice (C), infected mice at 24 h after infection (D), and infected mice at 48 h after infection (E ). Insets show microvascular endothelial cells and parenchymal cells. The scale bar denotes 100 mm. F and G, Expression of L-selectin and CXCL16 in the brains of uninfected control mice, infected wild-type mice, MyD88/ mice, and Toll-like receptor (TLR) 2/TLR4/ mice determined by protein array analysis (F ). Optical densities of CXCL16 expression are expressed in the percentage of positive controls (G).

Article Snippet: Slides were then incubated overnight at 4 C with a rat anti–mouse CXCL16 antibody (R&D Systems) or the appropriate isotype control immunoglobulin.

Techniques: In Vitro, In Vivo, Expressing, Control, Standard Deviation, Immunohistochemical staining, Staining, Infection, Protein Array

Journal: The Journal of infectious diseases

Article Title: CXCL16 contributes to neutrophil recruitment to cerebrospinal fluid in pneumococcal meningitis.

doi: 10.1086/656532

Figure Lengend Snippet: Figure 2. Functional analysis of CXCL16. A and B, Cerebrospinal fluid (CSF) pleocytosis (CSF white blood cells) (A) and bacterial cerebellar titers (expressed as colony-forming units, presented in log titers) (B) in mice infected with Streptococcus pneumoniae and treated intraperitoneally either with a monoclonal antibody to murine CXCL16 (CXCL16-Ab) or with an IgG2A isotype control (isotype) before (left) (7 animals per group) or 24 h after (right) (11 animals per group) infection. Evaluation was performed 24 h after administration of the antibody to each group. C–F, Fluoresence-activated cell sorter analyses on Hoxb8 neutrophils. C, Expression of the neutrophil marker Mac-1. D, Expression of the CXCL16 receptor CXCR6 in unstimulated neutrophils (31%). E, Expression of the CXCL16 receptor CXCR6 in neutrophils stimulated with tumor necrosis factor (TNF)–a (56%). F, Expression of the CXCL16 receptor CXCR6 in neutrophils stimulated with ethanol-killed Sreptococcus pneumoniae (EKP) (66%). G, Chemotactic activities of CXCL16 on Hoxb8 neutrophils using a microchemotaxis chamber. Cell migration is expressed as the mean number of leukocytes that migrated per field. MF, microscopic field; rCXCL16, diluted recombinant CXCL16. H, Expression of the typical neutrophil-attracting chemokines CXCL1 (KC), CXCL2 (MIP-2), and CXCL5 (LIX) in mouse brain homogenates, as determined using a mouse cytokine antibody array. Optical densities of all 3 chemokines in uninfected controls, infected isotype (isoAb)–, and CXCL16-antibody (CXCL16-Ab)–treated animals (4 per group, done twice) are expressed in a percentage of positive controls.

Article Snippet: Slides were then incubated overnight at 4 C with a rat anti–mouse CXCL16 antibody (R&D Systems) or the appropriate isotype control immunoglobulin.

Techniques: Functional Assay, Infection, Control, Expressing, Marker, Migration, Recombinant, Ab Array

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

Article Title: CXCR4 inhibition in human pancreatic and colorectal cancers induces an integrated immune response

doi: 10.1073/pnas.2013644117

Figure Lengend Snippet: The effect of CXCL12-stimulated CXCR4 on chemokine receptor-mediated migration of human immune cells. ( Left ) The coexpression of CXCR4 with ( A ) CXCR1, ( B ) CXCR3, ( C ) CXCR5, ( D ) CXCR6, and ( E ) CCR2 on human immune cell lines was evaluated by flow cytometry after staining with antibodies specific for the relevant chemokine receptors. Gray peaks indicate isotype controls. ( Center ) The effect of CXCL12-stimulation of CXCR4 on the chemotactic responses of A CXCR1-coexpressing Jurkat T lymphoblastoid cells to CXCL8, ( B ) CXCR3-coexpressing HSB2DP T lymphoblastoid cells to CXCL10, ( C ) CXCR5-coexpressing Raji B lymphoblastoid cells to CXCL13, ( D ) CXCR6-coexpressing Jurkat T lymphoblastoid cells to CXCL16, and ( E ) CCR2-coexpressing Molm13 monocytoid cells to CCL2 was assessed by including CXCL12 in the upper chamber (blue) and the other chemokines in the lower chamber (red) in the Boyden two-chamber assay. ( Right ) The chemotaxis assays were performed with the five cell lines when the placement of the chemokines in the Boyden chambers was reversed. Bar diagrams display mean and SEM (n = 3-4). Statistical analysis by Student’s t test: *** P < 0.001; **** P < 0.0001; ns, not significant.

Article Snippet: The concentration of each chemokine used is as following: CXCL8 (R&D Systems, 208-IL), 20 ng/mL; CXCL10 (R&D Systems, 266-IP), 1,000 ng/mL; CXCL13 (R&D Systems, 801-CX), 1,000 ng/mL; CXCL16 (R&D Systems, 976-CX), 50 ng/mL; CCL2 (R&D Systems, 279-MC), 200 ng/mL

Techniques: Migration, Flow Cytometry, Staining, Boyden Chamber Assay, Chemotaxis Assay

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a KEGG pathway enrichment of DEGs between FACS-sorted TNFR2 + and TNFR2 − CD4 + T cells from MPE ( n = 3). b , c Volcano plot ( b ) and heatmap ( c ) of DEGs related to the chemokine signaling pathway. d , e Flow cytometry histograms ( d ) and comparisons of chemokine receptor expression (CXCR6, CCR4 and CCR6) ( e ) on TNFR2 + T reg cells and TNFR2 − T reg cells ( n = 13). f Schematic of Transwell chemotaxis assay testing TNFR2 − T reg chemotaxis toward MPE supernatant (by Figdraw). g , h Transwell chemotaxis assay comparing TNFR2 + T reg frequencies between freshly isolated cells and cells that migrated toward MPE supernatant after 4 h incubation. i Concentrations of CXCL16, CCL17, CCL22 and CCL20 in MPE and PB were quantified using ELISA ( n = 16). j Schematic of chemotaxis assay to investigate the chemotatic axis to attract TNFR2 + T reg cells in MPE. k , l Flow cytometry histograms and comparisons of chemotaxis of TNFR2 + T reg cells in response to MPE in the presence of anti-CXCL16, anti-CCL17, anti-CCL22 or anti-CCL20 mAbs. Data shown in d , e , g – i , k and l are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using paired two-tailed Student’s t -test ( e and h ), Wilcoxon test ( i ) or one-way ANOVA ( l ). * P < 0.05, ** P < 0.01, **** P < 0.0001. ns not significant, mAbs monoclonal antibodies.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Flow Cytometry, Expressing, Chemotaxis Assay, Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Bioprocessing

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a Flow cytometry analysis comparing of CXCL16 expression on CD45 + and CD45 − cells from MPE ( n = 9). b CXCL16 expression on major cell populations of MPE: CAFs (CD45 − FAP + ), neutrophils (CD45 + CD11b + Ly6G + ), macrophages (CD45 + CD68 + ), T lymphocytes (CD45 + CD3 + ) and B lymphocytes (CD45 + CD19 + ) from MPE ( n = 8). c Schematic of CAF purification from MPE using MACS (by Figdraw). d Schematic of Transwell assay testing TNFR2 + T reg chemotaxis toward CXCL16-supplemented medium or CAF culture supernatant, with or without anti-CXCL16 mAbs (by Figdraw). e , f Representative flow cytometry plots ( e ) and comparisons ( f ) of TNFR2 + T reg frequencies recruited under different conditions ( n = 3). g Schematic diagram of the MPE mouse model. h Representative images showing mouse MPE and pleural cavity tumors. i – k Bioluminescence images depicting the growth ( n = 5) ( i ), MPE volume ( n = 5) ( j ) and Kaplan–Meier survival plot ( k ) of MPE mice ( n = 9–10 per group). l Concentrations of CXCL16 in MPE from mouse models ( n = 6). m , n Frequencies of TNFR2 + cells among T reg cells in murine MPE of each group. o , p Frequencies of CD8 + T cells from mouse MPE in each group. Data shown in a , b , e , f and h – p are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using paired two-tailed Student’s t -test ( a ), unpaired two-tailed Student’s t -test ( l ), one-way ANOVA ( b , f , j , n and p ) or log-rank test ( k ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns not significant, CS culture supernatant, MACS magnetic-activated cell sorting, α-CXCL16 CXCL16 neutralizing antibody.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Flow Cytometry, Expressing, Purification, Transwell Assay, Chemotaxis Assay, Two Tailed Test, FACS

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a GSVA of signaling pathways enriched in CXCL16 + CAFs compared with CXCL16 − CAFs. b , c Gene set enrichment analysis (GSEA) was performed on gene sets related to the hypoxia signaling pathway ( b ) and the glycolysis signaling pathway ( c ). A positive NES indicates higher expression in CXCL16 + CAFs. d , e Lactate levels in CAFs cultured under normoxia (21% oxygen) or hypoxia (1% oxygen) for 48 h or at indicated time points ( n = 3). f , g CXCL16 concentrations in CAFs supernatant under normoxia or hypoxia for 48 h or at indicated time points ( n = 3). h Western blot of CXCL16 expression in CAFs under hypoxia at designated times. i Schematic of metabolic modulators targeting glycolysis or lactate production. j , k Lactate levels ( j ) and CXCL16 concentrations ( k ) in CAFs treated with indicated glycolysis modulators for 48 h ( n = 3). l – n Dose-dependent effects of DCA ( l ), oxamate ( m ) or rotenone ( n ) on lactate and CXCL16 levels in CAFs supernatant. o Western blot analysis of CXCL16 expression in CAFs treated with the indicated concentrations of glycolysis modulators for 48 h. Data shown in d – h and j – o are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t -test ( d and f ) or one-way ANOVA ( e , g and j – n ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns not significant, NES normalized enrichment score, DMSO dimethylsulfoxide, DCA dichloroacetate.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Protein-Protein interactions, Expressing, Cell Culture, Western Blot, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a Western blot analysis of the indicated proteins in CAFs treated with increasing concentrations of exogenous lactate or glucose for 48 h. b CXCL16 concentrations in CAFs supernatant after exogenous lactate treatment. c , d Lactate ( c ) and CXCL16 ( d ) levels in CAFs supernatant after treatment with glucose for 48 h. e Western blot analysis of the indicated proteins in CAFs treated with DCA, oxamate or rotenone for 48 h. f Schematic of LDHA knockdown in CAFs. g Lactate in the culture supernatant of LDHA-knockdown CAFs. h Western blot analysis of the indicated proteins in LDHA-knockdown CAFs. i CXCL16 levels in the supernatant of LDHA-knockdown CAFs. j Western blot analysis of the indicated proteins in CAFs treated with increasing concentrations of β-alanine, lactylation modulator, for 48 h. k CXCL16 concentrations in CAFs supernatant after β-alanine treatment. Data shown in a – e and g – k are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using one-way ANOVA ( b – d , g , i and k ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns not significant, KD knockdown, sh-LDHA shRNA against LDHA, sh-NC negative control.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Western Blot, Knockdown, shRNA, Negative Control

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a IGV tracks presenting H3K18la enrichment at the CXCL16 gene locus in CAFs from MPE by CUT&Tag analysis. b ChIP–qPCR validation of H3K18la binding at the CXCL16 promoter. c – e Predicted FOXO3 binding sites at the CXCL16 promoter using JASPAR ( http://jaspar.genereg.net ). f ChIP–qPCR validation of FOXO3 binding at the CXCL16 gene promoter in CAFs. g Western blot analysis of FOXO3 and CXCL16 expression following FOXO3 knockdown or overexpression in CAFs. h , i CXCL16 levels in CAFs supernatants after FOXO3 knockdown ( h ) or overexpression ( i ). j Schematic of dual-luciferase reporter assay, drawn by Figdraw. k Luciferase assays assessing FOXO3-mediated regulation of CXCL16 transcription in CAFs. l , m CUT&Tag ( l ) and ChIP–qPCR ( m ) analyses showing H3K18la enrichment at the FOXO3 promoter. n Western blot analysis of the indicated proteins in CAFs treated with glucose (0–20 mm/l), DCA (0–20 mm/l), oxamate (0–20 mm/l), retenone (0–50 nm/l) or β-alanine (0–20 mm/l) for 48 h. o Western blot analysis of the indicated proteins in LDHA-knockdown CAFs. Data shown in b , f – i , k and m – o are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t -test ( h and i ) or one-way ANOVA ( b , f , k and m ). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns not significant, IGV Integrative Genomics Viewer, OE overexpression.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: ChIP-qPCR, Biomarker Discovery, Binding Assay, Western Blot, Expressing, Knockdown, Over Expression, Luciferase, Reporter Assay, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a Schematic illustrating the the generation of LDHA-knockout NIH/3T3 fibroblasts (LDHA −/− 3T3) using CRISPR–Cas9, drawn by Figdraw. b Western blot validation of LDHA knockout in LDHA −/− 3T3 fibroblasts. c , d Lactate ( c ) and CXCL16 ( d ) levels in the culture supernatant of LDHA −/− 3T3 fibroblasts. e Western blot analysis of the indicated proteins in LDHA −/− 3T3 fibroblasts cultured with or without glucose. f Schematic of MPE mouse model. g Representative images showing mouse MPE and thoracic tumors in each group. h – j, Bioluminescence images depicting the growth ( n = 5) ( h ), MPE volume ( n = 6) ( i ) and Kaplan–Meier survival curves (n = 10 per group) ( j ) of MPE-bearing mice. Data shown in c – e and g – j are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t -test ( c and d ), one-way ANOVA ( i ) and log-rank test ( j ). * P < 0.05, ** P < 0.01, *** P < 0.001, ns not significant, LDHA−/−3T3 LDHA-knockout NIH/3T3 fibroblasts.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Knock-Out, CRISPR, Western Blot, Biomarker Discovery, Cell Culture, Two Tailed Test

Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a , b Lactate ( a ) and CXCL16 ( b ) levels measured in MPE from mice treated with LDHA −/− 3T3 fibroblasts or control fibroblasts. c – j Flow cytometry plots ( c , e , g and i ) and comparisons ( d , f , h and j ) of the frequencies of TNFR2 + T reg cells ( c and d ), IFN-γ + CD8 + T cells ( e and f ), granzyme B + CD8 + T cells ( g and h ) and perforin + CD8 + T cells ( i and j ) in MPE. k Schematic diagram illustrating the proposed mechanism: CAFs in MPE undergo glycolysis, leading to elevated endogenous lactate levels. This increase in lactate induces H3K18 lactylation modification at the promoter regions of both the CXCL16 gene and its transcription factor FOXO3, thereby promoting CXCL16 expression. TNFR2 + T reg cells, which express high levels of CXCR6, the only known receptor for CXCL16, are efficiently recruited into MPE. This recruitment dampens the antitumor response generated by CD8 + T cells, leading to the immunosuppression and progression of MPE (by Figdraw). Data shown in a – j are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using unpaired two-tailed Student’s t -test ( a and b ) and one-way ANOVA ( d , f , h and j ). ** P < 0.01, *** P < 0.001, **** P < 0.0001. ns not significant.

Article Snippet: The membrane was blocked with 5% skim milk in Tris-buffered saline containing 0.05% Tween 20 (TBST) at room temperature for 2 h, followed by overnight incubation with primary antibodies at 4 °C, including anti-Pan-Kla antibody (#PTM-1401, PTM BIO), anti-H3K18la antibody (#PTM-1406RM, PTM BIO), anti-H3K9la antibody (#PTM-1419RM, PTM BIO), anti-H3K27la antibody (#PTM-1428, PTM BIO), anti-H4K12la antibody (#PTM-1411RM, PTM BIO), anti-H4K8la antibody (#PTM-1415RM, PTM BIO), anti-CXCL16 antibody (#60123-1-Ig, Proteintech), anti-FOXO3 antibody(#10849-1-AP, Proteintech), anti-GAPDH antibody (#60004-1-Ig, Proteintech) and anti-histone H3 antibody (#17168-1-AP, Proteintech).

Techniques: Control, Flow Cytometry, Modification, Expressing, Generated, Two Tailed Test

Journal: Hepatology Communications

Article Title: CXCR6 recruits Th2 cells and promotes liver fibrosis in schistosomiasis japonica

doi: 10.1097/HC9.0000000000000784

Figure Lengend Snippet: CXCL16 was primarily derived from macrophages in schistosome-associated fibrotic livers. (A) tSNE plot showing the distribution of different cell clusters. (B) Bubble plot of Cellchat analysis between different cell clusters in the CXCL signaling network. The closer to red, the greater the proportion of interaction. (C) Representative western blot images for CXCL16 in schistosome-associated fibrotic liver (SJ) and normal mouse liver (NC). (D) Violin plots of Cxcl16 in different cell clusters. (E) Proportions of different cell clusters in single-cell sequencing. (F–H) Expression of CXCL16 in macrophages. Gating strategy for macrophages (F4/80 + CD11b + CD45 + ). Statistical significance was determined using an unpaired t test, and the data are presented as mean ± SD (* p <0.05; ** p <0.01; *** p <0.001; **** p <0.0001). Abbreviation: MP, mononuclear phagocytes.

Article Snippet: Primary antibodies against α-SMA (1:1000, Zenbio) and CXCL16 (1:500, Proteintech) were incubated overnight at 4 °C, with GAPDH and β-tubulin serving as loading controls.

Techniques: Derivative Assay, Western Blot, Sequencing, Expressing

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 1. Expression of human CXCL16 in normal human skin. For chromogenic staining of CXCL16 and ADAM10 in healthy human skin, acetone-fixed sections were incubated with (a, d) rabbit anti-hCXCL16 (b) rabbit anti-ADAM10 or an irrelevant (c) rabbit IgG control antibody followed by incubation with POD-coupled goat anti-rabbit IgG antibody, enzymatic staining and counterstaining with hemalumn. In (a–c) epidermal skin and in (d) dermal skin containing a blood vessel (bottom right) is shown. Bar ¼ 50 mm.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Expressing, Staining, Incubation, Control

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 2. In situ expression and in vivo-release of murine CXCL16. (a) Paraformaldehyde-fixed fresh frozen sections of murine skin were incubated with radioactively labeled antisense cRNA probes (as) for mCXCL16 and sense- controls (s), respectively. Hybridized cRNA probes were subsequently visualized by autoradiography. Bar ¼ 20mm. (b) Wound fluid from injured mice was collected every 24hours over a period of 15 days, cleared by centrifugation and subsequently analyzed for the presence of CXCL16 by an ELISA specific for mCXCL16. Data are given as mean and SD (n ¼ 5). Statistically significant release of mCXCL16 (Po0.05) is indicated by asterisks.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: In Situ, Expressing, In Vivo, Incubation, Labeling, Autoradiography, Centrifugation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 3. Surface expression and release of CXCL16 by cultured keratinocytes. (a) HaCaT cells were harvested and analyzed for CXCL16 surface expression by flow cytometry using a purified rabbit anti-hCXCL16 antibody that was detected by a phycoerythrin-conjugated secondary antibody. The fluorescence signal of the cells stained for hCXCL16 is shown in comparison to that of unstained cells or cells receiving an isotype control antibody. (b) HaCaT cells were incubated in serum-free medium for different periods of time in the presence or absence of marimastat (5 mM). Subsequently, conditioned media were harvested and analyzed for released CXCL16 by a specific ELISA for hCXCL16. (c) PAM212 cells were incubated with a rat anti-mCXCL16 antibody, an isotype control antibody, or left unstained. After incubation with a FITC-conjugated secondary antibody cells were analyzed by flow cytometry. (d) PAM212 cells were incubated in serum-free medium for 4 hours in the presence or absence of marimastat (5 mM). Subsequently, released CXCL16 in the conditioned media was determined by a specific ELISA for mCXCL16. Data are representative for three experiments and shown as mean and SD (n ¼ 3). Asterisks indicate statistically significant differences (Po0.05) between inhibitor-treated cells and untreated cells.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Expressing, Cell Culture, Flow Cytometry, Purification, Fluorescence, Staining, Comparison, Control, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 4. Effect of ADAM10/ADAM17-inhibitors on CXCL16-release from cultured keratinocytes. (a) Cell lysate of cultured HaCaT cells was investigated for the presence of immature and processed forms of the metallproteinaseses ADAM10 and ADAM17. For Western blotting purified rabbit antibodies against the C-terminus of ADAM10 and ADAM17, respectively were used. (b–d) (b) HaCaT cells (c) human primary keratinocytes, (d) WT-ECV304 and CXCL16-ECV304 cells and were incubated for 4 hours in the presence or absence of 5 mM GI254023X or GW280264X. Subsequently, conditioned media were harvested and cell lysates were prepared. Released and cell-associated CXCL16 in media and lysates, respectively, was then quantified by ELISA. Statistically significant differences (Po0.05) between inhibitor-treated cells and untreated cells are indicated by asterisks.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Cell Culture, Western Blot, Purification, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 5. Effect of ADAM10/ADAM17 downregulation by siRNA on CXCL16-release from kreatinocytes. Cultured keratinocytes prepared from human foreskin were transfected with different heteroduplexed siRNA oligonucleotide constructs for downregulation of ADAM10 (A10-1, 2, 3) or ADAM17 (A17-1, 2, 3), an irrelevant siRNA control or without siRNA. At 48 hours after transfection mRNA and protein was extracted. (a and b) The mRNA expression level of ADAM10 and ADAM17 was determined by real-time RT-PCR and expressed in relation to that of glyceraldehyde-3-phosphate dehydrogenase. (c) Lysates of siRNA-transfected cells were analyzed for expression of the pro- and mature form of ADAM10 and controlled for b-actin content by Western blotting. (d and e) For CXCL16 release experiments siRNA-treated cells were washed and incubated for 4 hours in fresh medium. (c and e) Subsequently, cells were harvested for analysis of ADAM10 and ADAM17 mRNA expression and media were collected for quantification of released CXCL16 by ELISA. Data are representative for three experiments and shown as mean and SD (n ¼ 3). Asterisks indicate statistically significant differences (Po0.05) between cells receiving specific siRNA and control cells treated with irrelevant siRNA.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Cell Culture, Transfection, Construct, Control, Expressing, Quantitative RT-PCR, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

Journal: The Journal of investigative dermatology

Article Title: Constitutive expression and regulated release of the transmembrane chemokine CXCL16 in human and murine skin.

doi: 10.1038/sj.jid.5700751

Figure Lengend Snippet: Figure 8. Activity of keratinocyte-derived CXCL16 on CXCR6-transfected HEK293 cells. CXCR6-expressing HEK293 cells were treated with recombinant CXCL16 or concentrated HaCaT-conditioned (HaCaT-CM) medium for 30 minutes at either (a) 371C or (b) 41C. In a separate experiment recombinant CXCL16 or concentrated HaCaT-conditioned medium were preincubated with a neutralizing antibody to (c) CXCL16 or an (d) isotype control and then added to the CXCR6-expressing HEK293 cells for 30 minutes at 371C. Subsequently, cells were assayed for CXCR6 surface expression using a phycoerythrin-labeled mAb to CXCR6. As a control, WT-HEK293 cells expressing no CXCR6 were stained in parallel. The mean intensity of the fluorescence signal from CXCL16 or HaCaT-CM-treated cells was calculated as percentage of that from untreated cells and is shown as insets. Data are representative for three experiments.

Article Snippet: Recombinant human CXCL16 extracellular domain and chemokine domain, recombinant human IFN-g, recombinant human tumor necrosis factor-a, goat anti-human CXCL16 (hCXCL16) antibody, rat anti-murine CXCL16 (mCXCL16) mAb (clone 142417), phycoery- thrin-conjugated and unconjugated mouse anti-human CXCR6 (hCXCR6) mAb (clone 56811), and rat anti-murine CXCR6 (mCXCR6) mAb (clone 221002) were obtained from R&D Systems (Wiesbaden, Germany).

Techniques: Activity Assay, Derivative Assay, Transfection, Expressing, Recombinant, Control, Labeling, Staining, Fluorescence