ccl9  (R&D Systems)

Journal: The Journal of Clinical Investigation

Article Title: Inducible CCR2 + nonclassical monocytes mediate the regression of cancer metastasis

doi: 10.1172/JCI179527

Figure Lengend Snippet: ( A – E ) Detection of the uptake of B10F10-GFP by I-NCMs in vitro ( A – C ) and in vivo ( D and E ). Experimental design ( A ) and uptake of B16F10-GFP by RFP-labeled splenic ( B ) or blood ( C ) I-NCMs at 36 hours following coculture. The GFP signal in I-NCMs was detected by flow cytometry ( B ) or ICC ( C ). Monocytes and B16 cells were cocultured at a ratio of 5:1. White and yellow scale bars: 100 and 10 μm, respectively, in C . Experimental design ( D ) and FACS detection of uptake of B16F10-GFP materials by monocyte subsets in blood ( E , left) or lung (right) in Nr4a1 –/– mice after retro-orbital injection of MDP and B16F10-GFP injection. ( F – K ) I-NCMs recruit NK cells through CCL6 release to sites of B16F10 melanoma metastasis. ( F and G ) Adoptive transfer of I-NCMs ( F ) and MDP treatment ( G ) increased NK cells in the lungs of B16F10-bearing Nod2 –/– and Nr4a1 –/– mice, respectively. ( H ) MDP-triggered attenuation of B16F10 colonization in Nr4a1 –/– mice was inhibited by depletion of NK cells using NK1.1 antibody. ( I ) Bulk RNA-Seq data showing higher expression of Ccl6 and Ccl9 , but not other detected chemokine genes, in I-NCMs. ( J ) Anti-CCL6 antibody reduced NK cells and ( K ) suppressed the MDP-mediated attenuation of B16F10 colonization in Nr4a1 –/– mice. Data are presented as mean ± SEM; n = 4–10 in each group; * P < 0.05, ** P < 0.01, *** P < 0.001; 2-tailed t test in F and J , Kruskal-Wallis test in H , and 1-way ANOVA in G , I , and K .

Article Snippet: Anti–mouse CCL6/C10 antibody (AB-487-NA), anti-Mouse CCL9/10/MIP-1 gamma Antibody (AF463), and goat IgG isotype control (AB-108-C) were purchased from R&D Systems.

Techniques: In Vitro, In Vivo, Labeling, Flow Cytometry, Injection, Adoptive Transfer Assay, RNA Sequencing Assay, Expressing

Journal: Environmental disease

Article Title: Swine barn dust stimulates CCL9 expression in mouse monocytes through PKC-delta activation

doi: 10.4103/ed.ed_16_20

Figure Lengend Snippet: Hog barn dust extract increases CCL9 expression in RAW264.7. Cells were treated for 1, 3, 6, or 24 h with either media or hog barn dust extract at 1% v/v and measured for CCL9. Bars represent average of 3 replicates performed 3 times. Error bars represent standard error mean. 1 = significant between media treatments, P < 0.05 or lower; 2 = significant between hog barn dust extract treatments, P < 0.05 or lower; 3 = significant between media and hog barn dust extract treatments, P < 0.05 or lower

Article Snippet: The membrane was blocked with 5% milk in tris-buffered saline + 0.1% Tween20 (TBST) (0.1% Tween), followed by overnight incubation with 1:1000 dilution of rabbit anti-mouse CCL9 (500-P117, Peprotech, Rocky Hill, NJ) at 4°C.

Techniques: Expressing

Journal: Environmental disease

Article Title: Swine barn dust stimulates CCL9 expression in mouse monocytes through PKC-delta activation

doi: 10.4103/ed.ed_16_20

Figure Lengend Snippet: CCL9 is produced in response to lipopolysaccharide and peptidoglycan. RAW264.7 cells were treated 6 h with either media, lipopolysaccharide, peptidoglycan, hog barn dust extract (1% v/v), or hog barn dust extract that was boiled for 1 h or scrubbed with polymyxin B. Bars represent CCL9 protein average of 3 replicates performed 3 times. Error bars represent standard error mean. **P < 0.01, ****P < 0.0001

Article Snippet: The membrane was blocked with 5% milk in tris-buffered saline + 0.1% Tween20 (TBST) (0.1% Tween), followed by overnight incubation with 1:1000 dilution of rabbit anti-mouse CCL9 (500-P117, Peprotech, Rocky Hill, NJ) at 4°C.

Techniques: Produced

Journal: Environmental disease

Article Title: Swine barn dust stimulates CCL9 expression in mouse monocytes through PKC-delta activation

doi: 10.4103/ed.ed_16_20

Figure Lengend Snippet: Protein kinase C-δ inhibitor is involved in production of CCL9. RAW264.7 cells were treated 1 h prior to administration of hog barn dust extract with inhibitors to protein kinase C-α, δ, and ζ. hog barn dust extract was then administered and cells incubated an additional 6 h with inhibitors still present. Bars represent average of 3 replicates performed 3 times. Error bars represent standard error mean. *P < 0.01. NS = No significance

Article Snippet: The membrane was blocked with 5% milk in tris-buffered saline + 0.1% Tween20 (TBST) (0.1% Tween), followed by overnight incubation with 1:1000 dilution of rabbit anti-mouse CCL9 (500-P117, Peprotech, Rocky Hill, NJ) at 4°C.

Techniques: Incubation

Journal: Environmental disease

Article Title: Swine barn dust stimulates CCL9 expression in mouse monocytes through PKC-delta activation

doi: 10.4103/ed.ed_16_20

Figure Lengend Snippet: Protein kinase C-δ siRNA blocks CCL9 protein expression. siRNA containing either a nontargeting control (null) or protein kinase C-δ inhibiting sequence (protein kinase C -δ) was transfected into LA4 cells for 24 h prior to treatment of cells with either media or hog barn dust extract for 6 h. Release of CCL9 was measured. Bars represent average of 3 replicates performed 3 times. Error bars represent standard error mean. *P < 0.05, ****P < 0.0001

Article Snippet: The membrane was blocked with 5% milk in tris-buffered saline + 0.1% Tween20 (TBST) (0.1% Tween), followed by overnight incubation with 1:1000 dilution of rabbit anti-mouse CCL9 (500-P117, Peprotech, Rocky Hill, NJ) at 4°C.

Techniques: Expressing, Control, Sequencing, Transfection

Journal: Environmental disease

Article Title: Swine barn dust stimulates CCL9 expression in mouse monocytes through PKC-delta activation

doi: 10.4103/ed.ed_16_20

Figure Lengend Snippet: CCL9 inhibits stimulated keratinocyte-derived chemokine protein expression. Purified CCL9 was administered (20 ng/mL) to RAW264.7 cells concurrent with lipopolysaccharide, peptidoglycan, or hog barn dust extract treatment and incubated for 6 h. Bars represent average of 3 replicates performed 3 times. Error bars represent standard error mean. *P < 0.05; ****P < 0.0001

Article Snippet: The membrane was blocked with 5% milk in tris-buffered saline + 0.1% Tween20 (TBST) (0.1% Tween), followed by overnight incubation with 1:1000 dilution of rabbit anti-mouse CCL9 (500-P117, Peprotech, Rocky Hill, NJ) at 4°C.

Techniques: Derivative Assay, Expressing, Purification, Incubation

Journal: Brain sciences

Article Title: Pharmacological Modulation of the MIP-1 Family and Their Receptors Reduces Neuropathic Pain Symptoms and Influences Morphine Analgesia: Evidence from a Mouse Model.

doi: 10.3390/brainsci13040579

Figure Lengend Snippet: Figure 4. The effects of CCL9 neutralizing antibody administered intrathecally (i.t.) according to timeline (A), at doses of 0.5, 2, and 4 µg/5 µL on mechanical (B) and thermal (C) hypersensitivity, and the influence of a CCL9 neutralizing antibody at a dose of 2 µg/5 µL plus morphine 2.5 µg/5 µL on mechanical (E) and thermal (F) hypersensitivity, administered according to timeline (D), 7 days CCI in mice. The data are presented as the mean ± SEM (naive n = 5; CCI n = 5–8). The results were evaluated using one-way ANOVA followed by Bonferroni’s post hoc test for comparisons of selected pairs. * p < 0.05, ** p < 0.01, and *** p < 0.001 indicate significant differences vs. V-treated group at each of the investigated time points: 1, 4, and 24 h for (B,C) graphs; * p < 0.05, ** p < 0.01, and *** p < 0.001 indicate significant differences vs. V + W-treated group for (E,F) graphs; # p < 0.05 indicates significant differences between the V + M- and nAb + M-treated groups for (E,F) graphs; and & p < 0.05 indicates significant differences between the nAb + W- and nAb + M-treated groups. Abbreviations: V: vehicle (PBS); W: vehicle (aqua pro injectione); nAb: neutralizing antibody; M: morphine; CCI: chronic constriction injury of the sciatic nerve.

Article Snippet: Administration of CCL3 and CCL9 Neutralizing Antibodies A single i.t. administration of CCL3 nAb (AF-450-NA, Mouse CCL3/MIP-1 alpha Antibody, R&D Systems; Minneapolis, MI, USA) or CCL9 nAb (AF463, Mouse CCL9/10/MIP-1 gamma Antibody, R&D Systems) was administered to CCI mice at the dose of 0.5, 2, or 4 μg/5 μL on Day 7, when mechanical and thermal hypersensitivity were fully developed.

Techniques:

Journal: Brain sciences

Article Title: Pharmacological Modulation of the MIP-1 Family and Their Receptors Reduces Neuropathic Pain Symptoms and Influences Morphine Analgesia: Evidence from a Mouse Model.

doi: 10.3390/brainsci13040579

Figure Lengend Snippet: Scheme 1. Pharmacological modulation of chemokines from MIP-1 family (CCL3 and CCL9) via neutralizing antibodies and their receptors (CCR1 by J113863, CCR5 by TAK-220 or AZD -5672) reduces neuropathic pain symptoms and influences morphine analgesia—evidence from mice model evoked by chronic constriction injury of the sciatic nerve.

Article Snippet: Administration of CCL3 and CCL9 Neutralizing Antibodies A single i.t. administration of CCL3 nAb (AF-450-NA, Mouse CCL3/MIP-1 alpha Antibody, R&D Systems; Minneapolis, MI, USA) or CCL9 nAb (AF463, Mouse CCL9/10/MIP-1 gamma Antibody, R&D Systems) was administered to CCI mice at the dose of 0.5, 2, or 4 μg/5 μL on Day 7, when mechanical and thermal hypersensitivity were fully developed.

Techniques:

Journal: eLife

Article Title: Cytotoxic T cells swarm by homotypic chemokine signalling

doi: 10.7554/eLife.56554

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-CCL9/10 (monoclonal rat IgG1) , R & D Systems , Cat. #: MAB463 RRID: AB_2259783 , (5 μg/ml).

Techniques: In Vivo, Cell Culture, Transduction, Construct, Sequencing, Produced, Transfection, Expressing, Plasmid Preparation, Clone Assay, Control, Recombinant, In Vitro, Mass Spectrometry, Cell Isolation, Selection, Reverse Transcription, Sandwich ELISA, Cytometry, Software