goat anti mouse cxcl16 polyclonal antibody  (R&D Systems)

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a UMAP visualization of 398188 cells epidermal cells across control (CE) and Ube2l3 △Epi (UE) mice, classified into 6 immune/non-immune clusters via Louvain algorithm. Cell cluster: cyc-T, cycling T cell, Krt, keratinocytes, Mel, melanocytes, NK/T, natural kill T cells and T cells, SG, sebaceous gland. b Marker expression dot plot for the 6 cell populations. Dot size = expressing cell fraction; color intensity = mean scaled expression. c Compositional changes of epidermal cell states in CE vs. UE. d Predicted ligand-receptor interactions between clusters. Red arrow: upregulated Cxcl16 (myeloid)→ Cxcr6 (NK/T) axis in UE. e Signaling activity of CXCR6-CXCL16 and top ligand-receptor pairs across cell types. f Dot plot showing the differentially expressed Vγ2 and Vγ4 specified marker ( Tcrg-V4, 5830411N06Rik and Cd163l ) in T-cell subsets. g Flow cytometry: Increased IL-17A⁺CXCR6⁺CD3⁺ T cells in Ube2l3 △Epi mice epidermis. n = 6/group. h Flow cytometry plots and quantification of the percentage of Vγ2 and Vγ3 cells in γδT cells in Epi-Ctrl and Epi -Ube2l3 △Epi groups. Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6. i Flow cytometric analysis of the percentage of TNFa + and IL-17A + Vγ2 + cells in Vγ2 + γδT cells between Epi-Ctrl and Epi -Ube2l3 △Epi groups. Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6. j Schematic experimental design showing Ube2l3 △Epi mice treated with or without anti-Vγ2mAb (10ug/g/week) intraperitoneally in four weeks. k Representative phenotype of Ube2l3 △Epi mice treated with or without anti-Vγ2mAb before (Week 0) and after 4-week treatment (Week 4). Histological analysis of H&E staining was calculated and showed a decrease in Vγ2 antibody group. ( n = 5 per group). Immunofluorence analysis of Vγ2 (red) and CD3 (green) was showed. white triangle, double stained Vγ2 and CD3. l Proportion of L-17A + γδT cells in γδT in Epi-Ctrl, Epi -Ube2l3 △Epi and the epidermis of mouse Ube2l3 △Epi group treated with anti-Vγ2mAb (Epi- Ube2l3 △Epi - Vγ2Ab). Epi-Ctrl, n = 5, Epi -Ube2l3 △Epi , n = 6, Epi- Ube2l3 △Ep i- Vγ2Ab, n = 3. m FPKM of CXCR6 in GSE53552 (left, NL, n = 23, LS, n = 25, LS-treated, n = 20), GSE117239 (middle, NL, n = 34, LS, n = 34, LS-treated, n = 30), and GSE117239 (right, NL, n = 50, LS, n = 49, LS-treated, n = 45). n UMAP plots of CXCR6 expression in human lymphoid cells in NE and PE group. o Heatmap results showed the average expression of CXCR6 and other markers in 8 groups in human lymphoid cells. p Immunofluorescence staining of psoriatic skin for CD3 (green), CXCR6 (red), CD8 (purple), and DAPI (blue). White triangle indicated CXCR6 + and CD8 + T cells (Scale bar: 50 um). q Representative flow cytometric plots and percentage of IL-17A + CXCR6 + T cells in CD3 + T cells in Epi-HD ( n = 6) and Epi-PSO ( n = 3). The proportion of CD4 + and CD8 + in IL-17A + CXCR6 + T cells. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( g , h , i , k and q ), one-way ANOVA with Tukey’s multiple comparisons ( l , m ). The Figure j was created in BioRender. Chen, X. (2025) https://BioRender.com/854rwn9 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Control, Marker, Expressing, Activity Assay, Flow Cytometry, Staining, Immunofluorescence

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a The single cell profile of 28,725 Keratinocytes (Krt) about nine classes conserved epidermis across UE and CE, delineated by Louvain clustering. Basal, basal keratinocytes ( Krt14 and Krt5 ), spinous cell ( Krt1 and Krt10 ), granular cell ( Spink5 and Calm5 , Krt17 , Krt79 ), hair follicle stem cells (HFSC) ( Cd34 , Postn , Ptn and Fst ). b Proportional abundance of KC states in CE vs. UE. c Scatter plots defining KC clusters. d Top upregulated genes in krt subset in scRNA-seq of UE vs CEs. e Dot plot of cytokines/ chemokines ( Cxcl16 , Ccl20 , Il23a , Tnf and Il1b) compared in each group in Krt subset. red arrow is Prolif.cell group. f Flow cytometric plots and percentage of CXCL16 expression in CD45 - cells in Epi-Ctrl ( n = 6), Epi-IMQ ( n = 6) and Epi- Ube2l3 △Epi group ( n = 5). g The immunoblotting of UBE2L3, CXCL16 and β -actin in lysates, soluble CXCL16 (sCXCL16) in the supernatants (super.) of cultured primary mouse keratinocytes in control group (KC-Ctrl) and in Ube2l3 knockout group (KC- Ube2l3 △Epi ). h Quantification of ( g ). n = 3 biological replicates. i The immunoblotting results of UBE2L3, Pro-IL1β and β -actin in lysates, IL-1β p35 and IL-1β p17 in supernatants of KC-Ctrl and KC- Ube2l3 △Epi . j Quantification of ( i ). n = 3 biological replicates. k Mouse control keratinocytes were stimulated with IL-1β (100 ng/ml), IL-17A (100 ng/ml) for 24 hours. Immunoblotting of STAT3, pSTAT3, CXCL16 and β -actin was carried out. l Quantification of ( k ). ( n = 3 biological replicates). m The R&D Luminex assay showed that the protein level of CXCL16 was increased in Epi- Ube2l3 △Epi group. n = 5. n , o Flow cytometric plots and percentage of TNFα and CXCL16 in the epidermis of healthy donors (Epi-HD, n = 3) and psoriatic patients (Epi-PSO, n = 3). p , q Normal human epidermal keratinocytes (NHKEs) were stimulated with rhIL1β (100 ng/ml) and rhIL-17A(100 ng/ml) for 24 hours and the STAT3, pSTAT3 and CXCL16 in lysates were measured by western blot. ( n = 3 biological replicates). Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( h , j , m , and o ), one-way ANOVA with Tukey’s multiple comparisons ( m , q ).

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Expressing, Western Blot, Cell Culture, Control, Knock-Out, Luminex

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a UMAP of 1647 myeloid cells from control (Mouse-CE) and Ube2l3 △Epi (Mouse-UE) epidermis, clustered into 9 subsets: mast cells/basophils (MC_Bas), neutrophils (Neu), Langerhans cells (LC), macrophages (Mac), type 1 conventional DCs (cDC1), type 2 conventional DCs (cDC2). b Proportional abundance of myeloid subsets in CE vs. UE. c Expression of cytokines ( Il1b ) and chemokine ( Cxcl16 , Cxcl10 and Adam10 ) in nine defined classes. Red arrow: Cxcl16 in mDC_cDC2 cluster. d Pseoudotime analysis of Cxcl16 gene in nine cluster above in myeloid cells. Blue circles indicate the enrichment of Cxcl16 in mDC_cDC2 and Mac. e Flow cytometry and percentage of CXCL16 in mouse DC group in Epi-ctrl and Epi- Ube2l3 △Epi . Among CXCL16 + DC, the proportion of CD11b + DC (cDC2) and CD103 + DC (cDC1) in all CXCL16 + DC. n = 6. f Schematic strategy for generating Ube2l3 Epi△ - cDC -/- mice . g The phenotype of Ube2l3 Epi△ mice and Ube2l3 Epi△ - cDC -/- mice at week 0, 4, and 8. H&E staining (week 8). Epidermal thickness qualification in each group was calculated in Ube2l3 Epi△ mice ( n = 3) and Ube2l3 Epi△ - cDC -/- mice ( n = 4). Immunofluorescence of CD11C (red), K14 (green) and DAPI (blue), was done in two groups. scale bar= 50 um. h Flow cytometric plots and the percentage of IL17A + in CD3 + T, γδT and αβT in the epidermis of each group were showed. n = 3 per group. i UMAP visualization of 3237 human myeloid cells states found in human normal epidermis (NE) and human psoriatic epidermis (PE). pDC, Representative markers used in classifying mouse myeloid cells. pDC, plasmacytoid dendritic Cells. LC Langerhans cell cluster. Prolif.T, proliferated associated T cell. cDC2_mDC, type 2 DC and mature DC. Mac, macrophage. Pre-DC_cDC1, pre-DC and type 1 DC, B cell, B lymphocyte. j Heatmap of CXCL16 expression across human myeloid clusters. k Flow cytometry of IL-23p19 and CXCL16 expression in the epidermis of healthy donors (HD) and psoriatic patients (PSO). n = 3 per group. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( h , e , g , h , and k ). Figure f was created in BioRender. Chen, X. (2025) https://BioRender.com/riyjlc8 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Control, Expressing, Flow Cytometry, Staining, Immunofluorescence

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: a The percentage of Vγ2 + γδT in γδT in Der-Ctrl and Der- Ube2l3 △Epi group. b Schematic strategy for studying the migration of Vγ2 + γδT cells with or without Cxcl16 for 24 hours. c The migrated cell was viewed by immunofluorence staining with DAPI (blue), five independent views of lower chambers of insects were calculated between Der-Ctrl ( n = 5) and Der- Ube2l3 △Epi group ( n = 5). d Next, epidermal Vγ2 + γδT cells were sorted and stimulated with rmCXCL16 for 48 hours, and flow cytometry to be carried out to analyze the IL-17A secretion percentage. e Percentage of Vγ2 + IL-17A + T in CD3 + T in isotype and rmCXCL16 group. n = 3 per group. f , g Ube2l3 △Epi mice were treated with isotype antibodies or anti-CXCL16 mAb. H&E staining of back skin and ear skin was performed. h Epidermal single cells were stimulated with or without recombinant human CXCL16 and flow cytometric analysis of IL-17A was showed. i The percentage of IL-17A + CD3 + T /CD3 + Tcells, CD8 + IL-17A + /IL-17A + T cells, CD4 + IL-17A + /IL-17A + T cells in isotype cytokine and rhCXCL16 cytokine. j In wild-type (WT) mice, intradermal injection of recombinant mouse CXCL16 (rmCXCL16) and rmIL-23 every other day for 14 days. k H&E staining of and ear skin was performed. l Skin thickness measurements (using vernier calipers) of ear skin in control ear, rmCXCL16 ear, n = 5 per group, and control ear, rmIL-23 group, n = 3 per group. m – o Flow cytometry analysis of Vγ2 + γδT cell and Vγ3 + γδT cells in γδT, IL-17A in CD3 + T cells, and CXCR6 + IL-17A + in CD3 + T cells. n = 4 per group. Data are presented as mean ± SEM, and p -values were calculated by unpaired, two-tail student’s t -test ( a , c , e , g , l ), paired, two-tail student’s t -test ( i , k , m , n , o ). The Figure d , f , h , j was created in BioRender. Chen, X. (2025) https://BioRender.com/a73juv2 .

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Migration, Staining, Flow Cytometry, Recombinant, Injection, Control

Journal: Nature Communications

Article Title: Single cell transcriptomics of human psoriasis and epidermal specific Ube2l3 deficient mice highlight CXCL16/CXCR6 involvement in psoriasis development

doi: 10.1038/s41467-025-64106-6

Figure Lengend Snippet: A psoriasis-like lesion mouse model was generated by conditional knock out Ube2l3 in epidermis ( Ube2l3 △Epi ), which was compared with human psoriasis scRNA-seq data and facilitate cross-species comparisons of differentiation dynamics and ligand-receptor pathways in epidermis. In particular, IL-17A was regulated by CXCR6 + Vγ2 + γδT in mouse while CXCR6 + CD8 + T in human. Ube2l3 reduction in keratinocytes activated IL-1β and then promote CXCL16 expression through STAT3 signaling. CXCR6 + γδT17/ Tc17cells are prevalent within the epidermal immune microenvironment of psoriasis. Neutralization of CXCL16 inhibits the progression of psoriasis-like lesions in Ube2l3 △Epi mice. (Created in BioRender. Chen, X. (2025) https://BioRender.com/c9dlkqv ).

Article Snippet: The experimental design is shown in Fig. . Anti-CXCL16: Mice received subcutaneous injections (s.c.) of 4 μg/g/week goat anti-mouse CXCL16 polyclonal antibody (AF503, R&D Systems) or goat IgG isotype control.

Techniques: Generated, Knock-Out, Expressing, Neutralization

Journal: bioRxiv

Article Title: Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8 + T cells in triple-negative breast cancer

doi: 10.1101/2025.01.29.635356

Figure Lengend Snippet: (A) UMAP of total CD8 + T cells from E0771.lmb tumors. (B) Enrichment of T ex subset specific gene signatures from gp33-specific T cells from chronic LCMV infection model (clone 13) isolated on day 15 and 30 (Giles et al. ). (C) Dotplot showing expression of stem-like, effector, exhaustion, costimulatory, chemokine receptor, killer cell lectin-like receptor (KLR), transcription factors, interferon-stimulated genes (ISG) and proliferation-related genes across clusters. (D) UMAP CD8 + T cells split by treatment, isotype (left), anti-4-1BB (right). (E) Ratio of specific T ex subsets following anti-4-1BB treatment. (F) Schematic diagram depicting workflow for generating T ex subset specific anti-4-1BB augmented transcriptional signatures from E0771.lmb scRNA-seq dataset and applying signatures to BRCA patient specimen data from TCGA and METABRIC. (G) Total BRCA Relapse-free survival from METABRIC study (top) and Distant-Disease free relapse survival from TCGA (bottom) after separating high (3 rd quantile) and low (1 st quantile) gene signature expression groups. (H-I) Representative flow plots showing higher CXCR6 expression in T int relative to T prog and T term (left), gMFI of CXCR6 (middle), frequency of CXCR6 + (right), from models 4T1 (H), E0771.lmb (I). (J) Correlation between CXCR6 and TNFRSF9 expression in human TGCA BRCA tumor (left) and TCGA BRCA normal and breast GTEx samples (right). Spearman’s rank-correlation coefficient r and associated p -value are shown. Analysis done on GEPIA2 webtool. (K) Kaplan-Meier analysis of 4T1 wild-type or Cxcl16 sgRNA #1 treated with isotype or anti-4-1BB on days 13 and 15 post-tumor injection. Primary tumors were surgically resected on day 17 and mice underwent survival analysis. (L) Lung metastatic burden (mass) at endpoints. (in K-L, wild type (Isotype), n = 13-14; Cxcl16 sgRNA #1 (isotype), n = 15; Cxcl16 sgRNA #1 (α4-1BB), n = 13). P values were determined using log-rank (Mantel-Cox) test (G, K), one-way ANOVA followed by post-hoc Tukey’s multiple comparisons test (H, I).

Article Snippet: Membranes were probed with primary Polyclonal Goat IgG anti-mouse Cxcl16 (R&D Systems AF503) at 1:2,000 dilution from 0.2 μg/ml stock, and primary Rabbit anti-mouse β-actin (Cell Signaling Technology #4967) at 1:1,000 dilution from stock concentration overnight at 4 °C.

Techniques: Infection, Isolation, Expressing, Injection

Journal: bioRxiv

Article Title: Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8 + T cells in triple-negative breast cancer

doi: 10.1101/2025.01.29.635356

Figure Lengend Snippet: (A) GEPIA2 analysis comparing expression of CXCL16 across human breast cancer subtype from TCGA. (B) Sanger sequencing results showing successful knockout of Cxcl16 using gRNA targeting Exon 2. (C-E) Protein level verification of knockout using flow cytometry (C and D) and western blot (E). Treatment = 20ng/ml IFNγ + 20ng/ml TNFα + 1µM GI254023X (ADAM10 inhibitor). (F) Proliferation assay using Promega Realtime-Flo MT viability assay over a period of 48 hours (data pooled from 3 independent experiments). (G-J) Cxcl16 knockout reduces lung metastatic burden. (G) Experimental schematic. (H) Primary tumor mass at time of surgical resection. (I) Kaplan-Meier analysis. (J) Lung metastatic burden (mass) at endpoints. (in H, n = 10 per group; in I and J, wild-type, n = 8-9, Cxcl16 sgRNA #1, n = 9, Cxcl16 sgRNA #2, n = 5). P values were determined using two-tailed paired Student’s t -test (D), one-way ANOVA followed by post-hoc Dunnett’s multiple comparisons test (H), log-rank (Mantel-Cox) test (I).

Article Snippet: Membranes were probed with primary Polyclonal Goat IgG anti-mouse Cxcl16 (R&D Systems AF503) at 1:2,000 dilution from 0.2 μg/ml stock, and primary Rabbit anti-mouse β-actin (Cell Signaling Technology #4967) at 1:1,000 dilution from stock concentration overnight at 4 °C.

Techniques: Expressing, Sequencing, Knock-Out, Flow Cytometry, Western Blot, Proliferation Assay, Viability Assay, Two Tailed Test

Journal: bioRxiv

Article Title: Neoadjuvant anti-4-1BB confers protection against spontaneous metastasis through low-affinity intratumor CD8 + T cells in triple-negative breast cancer

doi: 10.1101/2025.01.29.635356

Figure Lengend Snippet: (A) Experimental schematic. (B) Primary tumor mass at time of surgical resection (left) and representation image (right). (C) Kaplan-Meier analysis of 4T1 Cxcl16 sgRNA #1 treated with isotype or anti-CD8β. Tumors were surgically resected on day 17 post-tumor inoculation and mice underwent survival analysis. (D) Lung metastatic burden (mass) at endpoints for mice in C. (E) Kaplan-Meier analysis of 4T1 Cxcl16 sgRNA #1 treated with anti-4-1BB in combination with or without anti-CD8β. Tumors were surgically resected on day 17 post-tumor inoculation and mice underwent survival analysis. (F) Lung metastatic burden (mass) at endpoints for mice in E. (wild type (Isotype), n = 14; Cxcl16 sgRNA #1 (isotype), n = 15; Cxcl16 sgRNA #1 (αCD8β), n = 9; Cxcl16 sgRNA #1 (α4-1BB), n = 13; Cxcl16 sgRNA #1 (α4-1BB + αCD8β), n = 12). Data are presented as mean ± SEM. P values were determined using one-way ANOVA followed by followed by post-hoc Šídák’s multiple comparisons test (B), log-rank (Mantel-Cox) test (C, E).

Article Snippet: Membranes were probed with primary Polyclonal Goat IgG anti-mouse Cxcl16 (R&D Systems AF503) at 1:2,000 dilution from 0.2 μg/ml stock, and primary Rabbit anti-mouse β-actin (Cell Signaling Technology #4967) at 1:1,000 dilution from stock concentration overnight at 4 °C.

Techniques:

Journal: Pharmaceutics

Article Title: Resveratrol Inhibited ADAM10 Mediated CXCL16-Cleavage and T-Cells Recruitment to Pancreatic β-Cells in Type 1 Diabetes Mellitus in Mice

doi: 10.3390/pharmaceutics14030594

Figure Lengend Snippet: ( A ) Double immunofluorescence staining for insulin (red), a pancreatic β cell marker, and CXCL16 (green) was performed; basal expression of pancreatic β cell CXCL16 in normal control and RES-treated mouse tissues. Increased CXCL16 expression in pancreatic β cells was confirmed by the co-localization of CXCL16 with insulin. Marked reduction of CXCL16 expression in mouse pancreatic β cells of RES-treated mice with STZ was seen; ( B ) Fluorescence intensity of CXCL16 protein expression for all groups was quantified and blotted; ( C ) Serum levels of CXCL16 using ELISA in all treated groups show similar findings of immunofluorescence. Data represent mean ± SEM: a difference is significant compared to control group; b difference is significant compared to RES group; c difference is significant compared to diabetic group at p < 0.05.

Article Snippet: Goat anti-rabbit polyclonal CXCL16 Ab was obtained from Peprotech (London, UK).

Techniques: Double Immunofluorescence Staining, Marker, Expressing, Control, Fluorescence, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Serum cytokine/chemokine array in patients with AP. Serum levels of six chemokines, among 40 cytokines/chemokines investigated, were significantly altered in MAP and SAP patients. Serum levels of CCL21, CCL13, and CCL15 in MAP were significantly lower in patients than in controls. Serum levels of MIF were significantly lower in SAP patients than in MAP patients. Serum levels of CCL27 were significantly lower in SAP patients than in control patients. Serum levels of CXCL16 were significantly higher in SAP patients than in control patients. When Bonferroni method was adopted to correct multiple testing problem, only CXCL16 level was revealed to have a significant difference. MAP, mild acute pancreatitis; SAP, severe acute pancreatitis. Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Control

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Cxcl16 expression in acute necrotizing pancreatitis. ( A ) Protocols of acute necrotizing pancreatitis by repeated injection of cerulein. Cerulein (100 ug/kg) was injected every 1 h 8 times on 2 consecutive days into C57BL/6 mice. Four to five mice were sacrificed at the indicated time points. ( B ) Serum amylase level and ( C ) pathology score of the respective time-points. ( D ) H&E sections in the pancreas of mice treated with acute necrotizing pancreatitis regimen at 0, 9, 24, and 33 h, respectively. The section at 33 h shows disrupted acinar architecture, vacuolization and necrosis of acinar cells, and inflammatory cell infiltration. ( E ) Pancreatic mRNA expressions of Cxcl16 , Tnfα , Il6 , and Cxcl2 were determined by quantitative RT-PCR analysis. *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Expressing, Injection, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Cxcl16 −/− mice were resistant to the development of necrotizing pancreatitis. Cerulein (100 ug/kg) was injected into Cxcl16 -intact ( WT ) and Cxcl16 -deficient ( Cxcl16 −/− ) mice (n = 5 in each group) every 1 h 8 times on 2 consecutive days as decribed in Fig. ( A ) Serum amylase levels and ( B ) pathology scores of WT and Cxcl16 −/− mice. ( C ) H&E sections from WT and Cxcl16 −/− mice at 24 h and 33 h. ( D ) Immunostained (Gr1, F4/80, and CD3) sections from WT and Cxcl16 −/− mice at 33 h. ( E ) Semiquantitative assessment of infiltartion of neutrophils, macrophages, and T cells. The number of neutrophils, macrophages, and T cells were counted in 5 high powered fields in each pancreas sections of WT and Cxcl16 −/− mice at 33 h. ( F ) MPO levels determined by ELISA in the pancreatic lysates of WT and Cxcl16 −/− mice at 33 h. *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Injection, Enzyme-linked Immunosorbent Assay

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Acinar cell expression of Cxcl16. Cerulein (100 µg/kg) was injected into WT and Cxcl16 −/− mice every 1 h 8 times on 2 consecutive days as described in Fig. . ( A ) Cxcl16 immunostaining of pancreatic frozen sections from WT mice (0 and 33 h) and Cxcl16 −/− mouse (33 h). ( B ) Dual immunofluorescence of Cxcl16 (Alexa Fluor 488) and amylase (Alexa Fluor 594), or Cxcl16 (Alexa Fluor 594) and F4/80 (Alexa Fluor 488). Most of Cxcl16-expressing on the cell surface was positive for cytoplasmic amylase expression. ( C ) Macrophage-depleted AP model (Left, protocol). Clodronate liposomes (100 mg/kg) were injected at 24 h before the first cerulein injection. F4/80 and Cxcl16 mRNA expression in the pancreas at 33 h, relative to those of control mice at 0 h, was evaluated at 33 h. *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Expressing, Injection, Immunostaining, Immunofluorescence, Liposomes, Control

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Induction of Ccl9 by Cxcl16 in necrotizing acute pancreatitis. ( A ) Cytokine/chemokine array using pancreatic lysates from C57BL/6 mice and Cxcl16 −/− mice treated with necrotizing pancreatitis regimen as described in Fig. . The relative expression of cytokines and chemokines is shown together with representative images. The data presented were obtained from WT and Cxcl16 −/− mice at 33 h, and WT on 0 h. ( B ) Ccl9 , Vcam-1 , and Ccl2 mRNA expression, relative to those of WT mice at 0 h, was assessed by qPCR in the pancreas of WT and Cxcl16 −/− mice at 33 h. ( C ) Ccl9 immunostaining of pancreatic frozen sections from WT mice at 0 h and 33 h, and Cxcl16 −/− mice at 33 h. *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Expressing, Immunostaining

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Expression of Ccl9 by pancreatic acinar cells. ( A ) Amylase secretion by the rat acinar cell line AR42J upon stimulation with various concentrations of cerulein. ( B ) Cxcl16 and Ccl9 mRNA expression by AR42J upon stimulation with cerulein. ( C ) Ccl9 mRNA expression of AR42J upon stimulation with cerulein (10 −10 M) in combination with various concentrations of recombinant Cxcl16 protein (left). Ccl9 mRNA expression of AR42J upon stimulation with cerulein (10 −7 M) in the presence of neutralizing anti-Cxcl16 Ab or control Ab (right). ( D ) Cxcl16 and Ccl9 mRNA expression by pancreatic acinar cells isolated from WT and Cxcl16 −/− mice upon stimulation with cerulein (10 −7 M). *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Expressing, Recombinant, Control, Isolation

Journal: Scientific Reports

Article Title: Chemokine CXCL16 mediates acinar cell necrosis in cerulein induced acute pancreatitis in mice

doi: 10.1038/s41598-018-27200-y

Figure Lengend Snippet: Therapeutic effects of neutralizing Cxcl16 Ab in the necrotizing pancreatitis model. ( A ) Injection protocol of anti-Cxcl16 Ab in necrotizing pancreatitis model. ( B ) Serum amylase level, ( C ) representative H&E sections, and pathology score of control Ab or anti-Cxcl16 Ab-treated mice at 39 h. ( D ) Gr1 immunostained sections and neutrophil counts of control Ab or anti-Cxcl16 Ab-treated mice at 39 h. ( E ) Pancreatic Ccl9 mRNA, pancreatic Ccl9 protein, and serum Ccl9 protein levels in control Ab or anti-Cxcl16 Ab-treated mice at 39 h. n = 5 in each group. *p < 0.05 Results were shown as mean ± SD.

Article Snippet: The sections were fixed with ice-cold methanol and immunostained using the following primary antibodies: goat polyclonal antibodies for Cxcl16 (R&D Systems, Minneapolis, MN) and Ccl9 (R&D Systems), a rabbit polyclonal antibody for amylase (Sigma-Aldrich).

Techniques: Injection, Control