Journal: CNS Neuroscience & Therapeutics

Article Title: Atractylenolide III ameliorates spinal cord injury in rats by modulating microglial/macrophage polarization

doi: 10.1111/cns.13839

Figure Lengend Snippet: Table of antibodies used

Article Snippet: CCR7 , Rabbit polyclonal , bs−1305R , AB_11056896 , PE , Bioss , 1 μg/test.

Techniques: Conjugation Assay, Concentration Assay

Journal: CNS Neuroscience & Therapeutics

Article Title: Atractylenolide III ameliorates spinal cord injury in rats by modulating microglial/macrophage polarization

doi: 10.1111/cns.13839

Figure Lengend Snippet: ATL‐III regulated the M1/M2 polarization of microglia/macrophages in rats after SCI. (A–H) Typical immunohistofluorescence pictures of CD68 (green), CCR7 (red in A–C), Arg‐1 (red in E–G), and nuclei (blue) in the spinal cords of sham, control, and ATL‐III rats. (D,H) The numbers of CD68 + CCR7 + (M1 cells) and CD68 + Arg‐1 + (M2 cells) cells. (I–K, M–O) Typical flow cytometry images of myeloid tissues from sham, control, and ATL‐III rats. (L,P) Proportions of CD68 + CCR7 + and CD68 + CD206 + cells. The data are presented as the mean ± SD ( n = 6). * p < 0.05, ** p < 0.01 compared with the control rats

Article Snippet: CCR7 , Rabbit polyclonal , bs−1305R , AB_11056896 , PE , Bioss , 1 μg/test.

Techniques: Immunohistofluorescence, Flow Cytometry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Co-expression of CCR7 and MUC1 in ESCC sample. a The expression of CCR7 and MUC1 in ESCC detected by IHC; b the immunoreactivity score of MUC1 in group with CCR7 positive expression and CCR7 negative expression group; c the 3-year regional recurrence curve of patients with CCR7 and MUC1 positive/negative expression; d the 5-year survival curve of patients with CCR7 and MUC1 positive/negative expression

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Clinical characteristics and its relationship with CCR7/ MUC1 expression

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Clinical characteristics and its relationship with CCR7 & MUC1 expression

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques:

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: CCL21 induced the up-regulation of MUC1 in ESCC cell lines. a , b the mRNA and protein expression level of MUC1 and CCR7 in ESCC cell lines; c : CCL21 induced the increasing MUC1 mRNA in ESCC cells. KYSE150, KYSE410, KYSE450 and Eca9706 were starved for 24 h and then treated with CCL21 at a concentration of 100 ng/ml for 12 h, then cells were harvested for the qRT-PCR; d , e CCL21 up regulated mRNA of MUC1 in a time and dose dependent way. Eca9706 and KYSE410 were starved for 24 h and then treated with CCL21 at a concentration of 0, 25, 50, 100, 200 ng/ml for 12 h or cells were treated with CCL21 at a concentration of 100 ng/ml for 0, 2, 6, 12 or 24 h. Then cells were harvested for qRT-PCR; f , g increasing of MUC1-C in KYSE410 and Eca9706 after treated with CCL21 at concentration of 0, 25,50, 100, 200 ng/ml for 24 h confirmed by immunoblotting; h , f blocking CCR7suppressed the up-regulation of MUC1 induced by CCL21. Eca9706 and KYSE410 were pretreated by CCR7 antibody (1 ug/mL) or IgG as control, then treated by 100 ng/ml for 24 h, then cells were harvested for detection of MUC1 by immunoblotting; j expression of MUC1 in KYSE410 and Eca9706 treated with PBS or CCL21(100 ng/mL) detected by immunofluorescence. Each data point represents the mean ± SD of three repeated experiments. * P < 0.05

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Expressing, Concentration Assay, Quantitative RT-PCR, Western Blot, Blocking Assay, Control, Immunofluorescence

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Silencing of MUC1 suppressed migration and invasion induced by CCL21. Cells were starved for 24 h and then seeding into the upper chamber, for the migration assay CCL21 was added into the lower chamber at a concentration of 200 ng/ml and incubated for 12 h; for the invasion assay the CCL21 was added into the upper chamber at a concentration of 200 ng/ml and incubated for 36 h. Cells on the lower surface of the membrane were counted in five randomly selected fields. a CCL21 promoted migration and invasion of KYSE410 and Eca9706 while blocking CCR7 could reverse migration and invasion induced by CCL21, and silencing MUC1 by siRNA targeted to MUC1 significantly suppressed the migration and invasion induced by CCL21; b Total cell numbers on the lower surface of the membrane counted in five randomly selected fields. Each data point represents the mean ± SD of 3 repeated experiments. * P < 0.05; Each datapoint represents the mean ± SD of three repeated experiments. * P < 0.05

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Migration, Concentration Assay, Incubation, Invasion Assay, Membrane, Blocking Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Activation of ERK1/2 was responsible for the up-regulation of MUC1 induced by CCL21-CCR7. a The activation of ERK1/2 and Akt pathway induced by CCL21, KYSE410 and Eca9706 cells were seeding into 6 well culture plate and starved overnight, then culture medium was replaced by serum free medium contained CCL21 (100 ng/ml) and incubated for 0, 15, 30, 45, 60 min. Then cells were harvested for immunoblotting; b Blocking CCR7 could suppress the activation of Akt and ERK1/2 pathway induced by CCL21 in KYSE410 and Eca9706; c , d Inhibiting activation of ERK1/2 but not Akt could suppress the up-regulation of MUC1-C protein. The starved KYSE410 and Eca9706 cells were pretreated by DMSO as controll, U0126 or MK2206 for 30 min, then cells were treated with PBS or CCL21 (100 ng/ml). For detecting p-ERK1/2, p-Akt and MUC1-C, cells were harvested after incubated with CCL21 for 15 min, 30 min and 24 h respectively; e Inhibiting ERK1/2 but not Akt could remarkably suppress the activity of MUC1 promoter, KYSE410 and Eca9706 cells transfected with MUC1-pGL2b plasmid were pretreated with DMSO as control, U0126 or MK2206 and then treated with PBS or CCL21 (100 ng/ml) for 12 h, then cells were harvested to detect the relative luciferase activity. Each datapoint represents the mean ± SD of three repeated experiments. * P < 0.05

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Activation Assay, Incubation, Western Blot, Blocking Assay, Activity Assay, Transfection, Plasmid Preparation, Control, Luciferase

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Phosphorylation of SP1 was responsible for the up-regulation of MUC1 induced by CCL21-CCR7. a , b Silencing Sp1 could remarkably suppressed the up-regulation of MUC1 in KYSE410 and Eca9706 induced by CCL21. The starved siSp1-Eca9706, siNC-Eca9706, siSp1-KYSE410 and siNC-KYSE410 were treated with CCL21(100 ng/mL) for 24 h, then cells were harvested for immunoblotting for MUC1-C; c Silencing Sp1/Mutant of Sp1 binding site at −99/−97 could remarkably suppressed MUC1 promoter activity induced by CCL21. siSp1-Eca9706, siNC-Eca9706, siSp1-KYSE410 and siNC-KYSE410 transfected by MUC1-pGL2b luciferase reporter plasmid/KYSE410 and Eca9706 cells transfected by MUC1-pGL2b or MUC1 mutant-pGL2b -firefly luciferase reporter plasmid were treated with PBS or CCL21(100 ng/mL) for 24 h, then cells were harvested for detecting the luciferase activity; d Increasing phosphorylation of Sp1 induced by CCL21. Starved KYSE410 and Eca9706 cells were treated with CCL21(100 ng/mL) for 0, 0.5, 1,2 and 6 h, then cells were harvested for the immunoblot of p-Sp1; e The expression of p-Sp1 in KYSE410 treated with PBS or CCL21(100 ng/ml) for 6 h detected by immunofluorescence; f Blocking CCR7 could suppress the phosphorylation of Sp1 induced by CCL21. The starved KYSE410 and Eca9706 cells pretreated with the CCR7 antibody or IgG as control, were treated with PBS or CCL21 for 6 h, then cells were harvested for the immunoblot of p-Sp1; g Inhibiting ERK1/2 suppressed phosphorylation of SP1 induced by CCL21. Starved KYSE410 and Eca9706 cells were pretreated with DMSO as control or U0126 for 30 min. After treated with PBS or CCL21(100 ng/mL) for 6 h, the cells were harvested for immunoblot; h Inhibiting ERK1/2 suppressed Sp1 binding to MUC1 promoter at −99/−97. Starved KYSE410 cells were pretreated with DMSO as control or U0126 for 0.5. After treated with PBS or CCL21(100 ng/mL) for 12 h, the cells were harvested for the ChIP assay. The targeted DNA was amplified using MUC1 primers with 40 cycles of PCR. Each datapoint represents the mean ± SD of three repeated experiments. * P < 0.05

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Phospho-proteomics, Western Blot, Mutagenesis, Binding Assay, Activity Assay, Transfection, Luciferase, Plasmid Preparation, Expressing, Immunofluorescence, Blocking Assay, Control, Amplification

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: CCL21-CCR7 promotes the lymph node metastasis of esophageal squamous cell carcinoma by up-regulating MUC1

doi: 10.1186/s13046-015-0268-9

Figure Lengend Snippet: Heterologous CCR7 promoted migration and invasion and up-regulated expression of MUC1 in KYSE150. a Heterologous expression of CCR7 up regulated the expression of MUC1 mRNA in KYSE150, KYSE150-CCR7 and KYSE150NC cells were starved and then treated with PBS or CCL21(0, 25, 50, 100, 200 ng/mL) for 12 h and then harvested for qRT-PCR and the result showed the remarkable up-regulation of MUC1 in KYSE150-CCR7 after treated with CCL21 compared to the KYSE150NC groups; b Heterologous expression of CCR7 up regulated the expression of MUC1-C protein in KYSE150, KYSE150-CCR7 and KYSE150NC cells were starved and then treated with PBS or CCL21(100 ng/mL) for 24 h and then harvested for immunoblot and the result showed the remarkable up-regulation of MUC1 in KYSE150-CCR7 after treated with CCL21 compared to the KYSE150NC groups; c Heterologous expression of CCR7 promoted migration and invasion induced by CCL21, the starved KYSE150-CCR7 and KYSE150NC cells were seeding into the upper chamber, for the migration assay CCL21 was added into the lower chamber at a concentration of 200 ng/ml and incubated for 12 h; for the invasion assay the CCL21 was added into the upper chamber and incubated for 36 h; d Total cell numbers on the lower surface of the membrane counted in five randomly selected fields. Each data point represents the mean ± SD of three repeated experiments. * P < 0.05

Article Snippet: Specific mouse polyclonal anti-MUC1 antibody (Abcam, UK) and specific rabbit polyclonal anti-CCR7 antibody (Abcam, UK) were used at a dilution of 1:100 and incubated at 4 °C overnight.

Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot, Concentration Assay, Incubation, Invasion Assay, Membrane

Journal: Acta biomaterialia

Article Title: Design principles for cytokine-neutralizing gels: Cross-linking effects

doi: 10.1016/j.actbio.2010.06.029

Figure Lengend Snippet: Quantification of immunohistochemical staining of tissue sites treated as noted. Bars shaded with vertical lines were stained for the pan-macrophage marker CD68, bars with diagonal shading were stained for the macrophage M2 phenotype marker CD163 and bars shaded solid black were stained for the macrophage M1 marker CCR7. Select p-values are written above the double-headed arrows connecting results that have significantly different values.

Article Snippet: The secondary antibodies used were biotinylated horse anti-mouse IgG (Vector, CD68 and CD163) at a dilution of 1:50 and goat anti-rabbit IgG (CCR7, Vector) at a dilution of 1:100.

Techniques: Immunohistochemical staining, Staining, Marker

Journal: Journal of Clinical and Diagnostic Research : JCDR

Article Title: Evaluation of Tumour Associated Macrophages and Angiogenesis in Ameloblastoma

doi: 10.7860/JCDR/2017/28067.10568

Figure Lengend Snippet: CCR7+ immune cells (marked by red arrows) and CCR7 negative immune cells (marked by black arrows) adjacent to islands of odontogenic epithelium X400.

Article Snippet: Materials and Methods Forty-six Formalin Fixed Paraffin Embedded (FFPE) blocks of ameloblastoma were processed for Abcam Mouse monoclonal Anti-CCR7 antibody, Abcam Rabbit polyclonal Anti-CD206 antibody and Dako Mouse monoclonal Anti-CD34 antibody QBEnd-10.

Techniques:

Journal: Journal of Clinical and Diagnostic Research : JCDR

Article Title: Evaluation of Tumour Associated Macrophages and Angiogenesis in Ameloblastoma

doi: 10.7860/JCDR/2017/28067.10568

Figure Lengend Snippet: Relative topographical expression of TAMs and MVD in ameloblastoma.

Article Snippet: Materials and Methods Forty-six Formalin Fixed Paraffin Embedded (FFPE) blocks of ameloblastoma were processed for Abcam Mouse monoclonal Anti-CCR7 antibody, Abcam Rabbit polyclonal Anti-CD206 antibody and Dako Mouse monoclonal Anti-CD34 antibody QBEnd-10.

Techniques: Expressing

Journal: Journal of biomedical materials research. Part A

Article Title: Evaluation of the tissue response to alginate encapsulated islets in an omentum pouch model

doi: 10.1002/jbm.a.35769

Figure Lengend Snippet: Immunohistochemical staining around alginate microbeads. Spleen was used as a control for markers CD68 (A), CD163 (D), CCR7 (G). CD68+ stain is observed (B, C)as well as CCR7+ (H,I). CD163+ staining was not observed (E, F). (C, F, I) higher resolution images of the inset area.

Article Snippet: The secondary antibody was added Biotinylated anti-mouse IgG (Vector, BA-2001, for CD68 and CD163) and Bio-tinylated anti-rabbit IgG (Vector, BA-1000, for CCR7) at a dilution of 1:200 for all antibodies.

Techniques: Immunohistochemical staining, Staining

Journal: Journal of biomedical materials research. Part A

Article Title: Evaluation of the tissue response to alginate encapsulated islets in an omentum pouch model

doi: 10.1002/jbm.a.35769

Figure Lengend Snippet: Masson’s Trichrome (A, B) indicates the presence of a chronic inflammatory response (red color) around both stable and failed alginate microbeads with islets and the presence of collagen for a small subset of alginate microbeads without islets. CCR7 stain indicates the presence of inflammatory macrophages (M1) in stable and failed alginate microbeads (C,D) while CD163 pro-healing macrophage phenotype (M2) was not observed in the groups with islets (E), the presence of pro-healing macrophage (CD163+) is observed in stable and failed alginate microbeads without islets (F). Arrows indicate macrophage presence.

Article Snippet: The secondary antibody was added Biotinylated anti-mouse IgG (Vector, BA-2001, for CD68 and CD163) and Bio-tinylated anti-rabbit IgG (Vector, BA-1000, for CCR7) at a dilution of 1:200 for all antibodies.

Techniques: Staining