goat igg anti mouse ccl21 antibody  (R&D Systems)

Journal: Scientific Reports

Article Title: Cholinergic anti-inflammatory pathway ameliorates murine experimental Th2-type colitis by suppressing the migration of plasmacytoid dendritic cells

doi: 10.1038/s41598-021-04154-2

Figure Lengend Snippet: The inhibitory effect of nicotine on BMpDC migration was evaluated with a chemotaxis assay. ( A ) BMpDCs matured by treatment with CpG oligodeoxynucleotides chronologically migrated in response to an established concentration gradient of CCL21 (a representative experiment is shown). ( B ) The number of migrating BMpDCs was dose-dependently inhibited by the addition of nicotine (1–100 μM) to the chemotaxis assay medium ( n = 3–7). ( C , D ) The velocity and directionality of BMpDC migrating in response to CCL21 were calculated. The velocity and directionality of BMpDC migration were significantly inhibited by the addition of nicotine (10 µM) to the chemotaxis assay medium, and pretreatment with MLA blocked the inhibitory effect of nicotine ( n = 20 for each group, ** P < 0.01 vs nicotine). Data are represented as the mean value ± SEM. P values were calculated using one-way ANOVA with Dunnett’s multiple comparison test ( C , D ).

Article Snippet: CCL21, a goat IgG anti-mouse CCL21 antibody, AZ-10417808, GM-CSF and FLT3 ligand were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, Chemotaxis Assay, Concentration Assay, Comparison

Journal: Scientific Reports

Article Title: Cholinergic anti-inflammatory pathway ameliorates murine experimental Th2-type colitis by suppressing the migration of plasmacytoid dendritic cells

doi: 10.1038/s41598-021-04154-2

Figure Lengend Snippet: The expression of active Rac 1 (Rac1-GTP) and total Rac 1 in BMpDCs was measured by western blotting. The ratio of active Rac 1 to total Rac 1 was significantly increased by stimulation with CCL21, and the increase in the ratio induced by CCL21 was inhibited by pretreatment with nicotine (10 µM) (a representative band pattern is shown; n = 4 for each group, * P < 0.05 vs CCL21). The full-length blots are shown in supplementary Fig. . Data are represented as the mean value ± SEM. P values were calculated using one-way ANOVA with Dunnett’s multiple comparison test.

Article Snippet: CCL21, a goat IgG anti-mouse CCL21 antibody, AZ-10417808, GM-CSF and FLT3 ligand were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Comparison

Journal: Scientific Reports

Article Title: Cholinergic anti-inflammatory pathway ameliorates murine experimental Th2-type colitis by suppressing the migration of plasmacytoid dendritic cells

doi: 10.1038/s41598-021-04154-2

Figure Lengend Snippet: The proportions of pDCs in the colonic mucosa and MLN of OXZ mice were analyzed. ( A , B ) The proportions of pDCs in the MLN and LPMCs of the middle colon in normal mice, OXZ mice, and OXZ mice treated with nicotine were compared. The frequency of pDCs in the MLN of OXZ mice was significantly decreased compared with that in the MLN of normal mice. The frequency of pDCs in LPMCs from OXZ mice was increased compared with that in the LPMCs from normal mice. The frequencies of pDCs in the MLN and LPMCs of OXZ mice treated with nicotine were equivalent to the corresponding frequencies of OXZ mice ( A : a representative experiment is shown, B : n = 4–6, ** P < 0.01 vs OXZ, n.s.: not significant). ( C ) The localization of CCL21 in OXZ mice was investigated by immunohistochemistry. CCL21 was mainly expressed around the T cell zone in ILFs (a representative result is shown). The scale bars represent 50 μm. Data are represented as the mean value ± SEM. P values were calculated using one-way ANOVA with Dunnett’s multiple comparison test ( B ).

Article Snippet: CCL21, a goat IgG anti-mouse CCL21 antibody, AZ-10417808, GM-CSF and FLT3 ligand were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Immunohistochemistry, Comparison

Journal: Scientific Reports

Article Title: Cholinergic anti-inflammatory pathway ameliorates murine experimental Th2-type colitis by suppressing the migration of plasmacytoid dendritic cells

doi: 10.1038/s41598-021-04154-2

Figure Lengend Snippet: Graphical abstract. The cholinergic anti-inflammatory pathway (nicotine and vagus nerve stimulation) ameliorated Th2-type OXZ colitis through α7nAChRs on pDCs, which was attributed to the suppression of pDC migration toward CCL21 in ILFs of the colonic mucosa of OXZ mice by α7nAChR-mediated JAK2-STAT3 activation, subsequent nonapoptotic caspase-3 activation and eventual Rac 1 inactivation.

Article Snippet: CCL21, a goat IgG anti-mouse CCL21 antibody, AZ-10417808, GM-CSF and FLT3 ligand were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Migration, Activation Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Suppression of plasmacytoid dendritic cell migration to colonic isolated lymphoid follicles abrogates the development of colitis.

doi: 10.1016/j.biopha.2021.111881

Figure Lengend Snippet: Fig. 4. Effects of 1 μM As-IV or Oxy on RAC1 activation. RAC1 activation was detected by evaluating the expression of the GTP-RAC1 protein by using a western blot analysis kit. A representative image and the quantification of the band intensity for GTP-RAC1 relative to that of total RAC1 are shown in A-C. (A) Representative western blot analysis of GTP-RAC1 in BMpDCs induced with CCL21 and quantification of the band intensity are shown (5 independent experiments). Data are expressed as the mean ± SE (*p < 0.05, n = 5). Representative western blot analysis of GTP-RAC1 in CCL21-induced migrated BMpDCs following treatment with As- IV (B) or Oxy (C) and the quantification of band intensity are shown (7 independent experiments). Data are expressed as the mean ± SE (*p < 0.05, n = 7). Microarray analysis of BMpDCs induced with a CCL21 gradient following treatment with As-IV or Oxy was performed by using the Clariom S Array Mouse. The differential expression of genes in BMpDCs induced with a CCL21 gradient following treatment with vehicle (control), As-IV or Oxy is displayed in the heatmap (D). The red and blue colors indicated the up-regulation normalized intensity values (log2) and down-regulation normalized intensity values (log2) of each RNA in each sample.

Article Snippet: 30 μm sections cut by using a cryostat (Leica, Nussloch, Germany) were soaked in 0.3% Triton X (Sigma, Missouri, USA) for 2 h and 2% Block Ace (DS Pharma Biomedical, Osaka, Y. Zhang et al. Biomedicine & Pharmacotherapy 141 (2021) 111881 Japan) for 1 h. Then, the colon sections were stained with the primary antibodies rat IgG anti-mouse B220 (1:200, BioLegend, San Diego, CA, USA), hamster IgG anti-mouse CD11c (1:100, BioLegend) and goat IgG anti-mouse CCL21 (1:200, R&D Systems).

Techniques: Activation Assay, Expressing, Western Blot, Microarray, Quantitative Proteomics, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Suppression of plasmacytoid dendritic cell migration to colonic isolated lymphoid follicles abrogates the development of colitis.

doi: 10.1016/j.biopha.2021.111881

Figure Lengend Snippet: Fig. 6. Effects of As-IV or Oxy on the distribution of CCL21 in the DSS-induced colitis model. The distribution of CCL21 in the colonic ILFs of DSS-induced colitis mice treated with saline (A), As-IV (B) or Oxy (C) was identified by immunohistochemical staining of the colon. Immunohistochemical staining was performed on samples from 3 mice/group, and representative images are presented.

Article Snippet: 30 μm sections cut by using a cryostat (Leica, Nussloch, Germany) were soaked in 0.3% Triton X (Sigma, Missouri, USA) for 2 h and 2% Block Ace (DS Pharma Biomedical, Osaka, Y. Zhang et al. Biomedicine & Pharmacotherapy 141 (2021) 111881 Japan) for 1 h. Then, the colon sections were stained with the primary antibodies rat IgG anti-mouse B220 (1:200, BioLegend, San Diego, CA, USA), hamster IgG anti-mouse CD11c (1:100, BioLegend) and goat IgG anti-mouse CCL21 (1:200, R&D Systems).

Techniques: Saline, Immunohistochemical staining, Staining

Journal: Methods in Molecular Biology

Article Title: Biolistic DNA Delivery

doi: 10.1007/978-1-62703-110-3

Figure Lengend Snippet: Fig. 1. Epidermal expression of transgenic CCL21 in the skin upon gene gun-mediated administration of pVR1012-mCCL21-IRES-EGFP in vivo and its subsequent drainage into PLNs. ( a ) Expression of EGFP is restricted to the epidermis, and no fl uorescence is detected after application of the pVR1012-mock vector. Skin sections were obtained 24 h after pDNA administration. ( b ) Day-1 cryosections were immunostained using a polyclonal goat anti- mouse CCL21 Ab. Hematoxylin counterstaining. Note that CCL21 immunoreactivity in the epidermis is similar to the EGFP expression pattern. In addition, CCL21 immunoreactivity can be observed in the dermis, most probably representing CCL21 secreted from the epidermis into the dermis. No immunoreactivity after application of pVR1012-mock vector. ( c ) In vitro cultured whole single-cell suspension of skin 3 days after pVR1012-mCCL21-IRES-EGFP application. Secretion of mCCL21 as measured by mCCL21 ELISA in supernatants. Adapted with modi fi cation and permission from the Journal of Investigative Dermatology.

Article Snippet: Mouse CCL21/6Ckine DuoSet kit (DY457, R&D Systems, other ELISA kits for respective chemokines can be also used on below principle): (a) Capture Antibody: 720 m g/mL of polyclonal goat anti- mouse CCL21 IgG antibody (AF457, R&D Systems) reconstituted with PBS.

Techniques: Expressing, Transgenic Assay, In Vivo, Plasmid Preparation, In Vitro, Cell Culture, Suspension, Enzyme-linked Immunosorbent Assay