rabbit anti ccr8  (Boster Bio)

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Correlation of CCR8 expression with the migration and differentiation of hepatic macrophages during liver injury. (A) Representative images (scale bar = 100 μm) of CCR8-stained liver sections from healthy mice, n = 10 and CCl4 mice, n = 12. (B) Quantitative analysis of protein expression and relative mRNA expression of CCR8 in the liver tissues from healthy mice ( n = 10) and CCl4 mice ( n = 12). (C) Representative composite images (scale bar = 100 μm) of liver sections from healthy mice, n = 10 and CCl4 mice, n = 12 stained with CCR8 (red), F4/80 or CD11b (green), and DAPI (nuclear staining, blue). (D) Correlation plot between Ccr8 mRNA expression and Adgre1 (F4/80) mRNA expression. (E) Correlation plot between Ccr8 mRNA expression and% CD11b ++ F4/80 + MoMFs population. Results are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01; P denotes statistical significance, and r represents Pearson correlation coefficient.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques: Expressing, Migration, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: 3D structures and docking of peptides: AP8i (orange), AP8ii (yellow), AP8iv (red) and small molecule R243 (magenta), and their respective 3D docking visualization at the active site of CCR8 (Homology build model).

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: 3D and 2D docking poses of (A) AP8ii and (B) R243 at the CCR8 active site based on the homology build model.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Molecular dynamic simulation studies of AP8ii at the active site of CCR8 (Homology Build Model). (A) RMSD graph showing AP8ii interaction at the CCR8 active site (Homology Build Model) for 100 ns. (B) 2D MD simulation pose of AP8ii at the active site of CCR8 (Homology build model). (C) Interactions (fraction) of peptide AP8ii at the CCR8 active site via MD simulation at 100 ns showing the type of Protein–Ligand contacts.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Molecular dynamic simulation studies of R243 at the active site of CCR8 (Homology Build Model). (A) RMSD graph showing R243 interaction at the CCR8 active site (Homology Build Model) for 100 ns. (B) Interactions (fraction) of R243 at the CCR8 active site via MD simulation at 100 ns showing the type of Protein–Ligand contacts. Insert: 2D MD simulation pose of R243 at the active site of CCR8 (Homology build model).

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: In vitro binding analysis of CCR8 antagonizing peptides on LPS/IFN γ -activated RAW264.7 macrophages. Representative images and quantitative analysis of AP8i, AP8ii, AP8iii, and AP8iv binding to LPS/IFN γ -activated RAW264.7 macrophages after 4 h of incubation at room temperature. The composite and zoomed in image (indicated with the dotted white box) showing DAPI-stained cells (nuclei are stained blue with DAPI) and RhB-labeled peptides (in green). Results from 4 independent experiments (performed in triplicates) are presented as mean ± SEM. ∗∗∗∗ P < 0.001; One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques: In Vitro, Binding Assay, Incubation, Staining, Labeling

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: AP8ii–CCR8 interaction analysis. Representative composite and zoomed in images (indicated with the dotted white box) of the composite images of pre-incubation (A) or co-incubation (B) studies performed on LPS/IFN γ -activated RAW264.7 macrophages. RhB-labelled AP8ii (10 μmol/L) that was either co-incubated with R243 (10 μmol/L), CCL1 (100 ng/mL), or the anti-CCR8 antibody (5.0 μg/mL) for 4 h or R243 (10 μmol/L), CCL1 (100 ng/mL), or the anti-CCR8 antibody (5.0 μg/mL) was preincubated for 2 h followed by incubation with AP8ii (10 μmol/L) for 4 h. (C) Surface Plasmon Resonance analysis using the CellVysion SPR imaging system. The first two columns show the peptide interaction with LPS/IFN γ -activated RAW264.7 and THP-1 macrophages, while the last two columns show peptide binding following anti-CCR8 antibody blocking. Results from 6 independent experiments (performed in duplicates) are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01; One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques: Incubation, SPR Assay, Imaging, Binding Assay, Blocking Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: AP8ii inhibits CCL1-induced migration and LPS/IFN γ -induced activation of macrophages in vitro . (A) Schematic representation of CCL1-induced migration assay; macrophages/monocytes are seeded in the Transwell insert with or without AP8ii, R243 or DMSO (10 μmol/L), and CCL1 (10 ng/mL) is added in the lower well. (B) Representative pictures of migrated nuclei (DAPI)-stained RAW macrophages. (C) Analyzed results of the migration assay performed on mouse RAW macrophages ( n = 3), BMDMs ( n = 3) and human THP1 monocytes ( n = 6). (D) Schematic showing the experimental setup for the experiment. (E) Graphs showing the gene expression analysis of Inos , Il6 , and Ccr8 performed on mouse RAW264.7 macrophages. The cells were activated with 100 ng/mL LPS + 10 ng/mL IFN γ with and without 10 μmol/L of CCR8 antagonizing peptide (AP8ii) or CCR8 antagonist (R243) with anti-CCR8 antibodies (CCR8AB) or 10 ng/mL IL4 and 10 ng/mL IL13 for 24 h followed by gene expression analysis. All results are presented as mean ± SEM from three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Thereafter, sections were rehydrated in PBS and incubated with the primary antibodies: rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals); rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or goat anti-mouse collagen-I (Cat. No. 1310-01, Southern Biotech, Birmingham, USA) at 4 °C overnight.

Techniques: Migration, Activation Assay, In Vitro, Staining, Gene Expression

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Correlation of CCR8 expression with the migration and differentiation of hepatic macrophages during liver injury. (A) Representative images (scale bar = 100 μm) of CCR8-stained liver sections from healthy mice, n = 10 and CCl4 mice, n = 12. (B) Quantitative analysis of protein expression and relative mRNA expression of CCR8 in the liver tissues from healthy mice ( n = 10) and CCl4 mice ( n = 12). (C) Representative composite images (scale bar = 100 μm) of liver sections from healthy mice, n = 10 and CCl4 mice, n = 12 stained with CCR8 (red), F4/80 or CD11b (green), and DAPI (nuclear staining, blue). (D) Correlation plot between Ccr8 mRNA expression and Adgre1 (F4/80) mRNA expression. (E) Correlation plot between Ccr8 mRNA expression and% CD11b ++ F4/80 + MoMFs population. Results are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01; P denotes statistical significance, and r represents Pearson correlation coefficient.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques: Expressing, Migration, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: 3D structures and docking of peptides: AP8i (orange), AP8ii (yellow), AP8iv (red) and small molecule R243 (magenta), and their respective 3D docking visualization at the active site of CCR8 (Homology build model).

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: 3D and 2D docking poses of (A) AP8ii and (B) R243 at the CCR8 active site based on the homology build model.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Molecular dynamic simulation studies of AP8ii at the active site of CCR8 (Homology Build Model). (A) RMSD graph showing AP8ii interaction at the CCR8 active site (Homology Build Model) for 100 ns. (B) 2D MD simulation pose of AP8ii at the active site of CCR8 (Homology build model). (C) Interactions (fraction) of peptide AP8ii at the CCR8 active site via MD simulation at 100 ns showing the type of Protein–Ligand contacts.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: Molecular dynamic simulation studies of R243 at the active site of CCR8 (Homology Build Model). (A) RMSD graph showing R243 interaction at the CCR8 active site (Homology Build Model) for 100 ns. (B) Interactions (fraction) of R243 at the CCR8 active site via MD simulation at 100 ns showing the type of Protein–Ligand contacts. Insert: 2D MD simulation pose of R243 at the active site of CCR8 (Homology build model).

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: In vitro binding analysis of CCR8 antagonizing peptides on LPS/IFN γ -activated RAW264.7 macrophages. Representative images and quantitative analysis of AP8i, AP8ii, AP8iii, and AP8iv binding to LPS/IFN γ -activated RAW264.7 macrophages after 4 h of incubation at room temperature. The composite and zoomed in image (indicated with the dotted white box) showing DAPI-stained cells (nuclei are stained blue with DAPI) and RhB-labeled peptides (in green). Results from 4 independent experiments (performed in triplicates) are presented as mean ± SEM. ∗∗∗∗ P < 0.001; One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques: In Vitro, Binding Assay, Incubation, Staining, Labeling

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: AP8ii–CCR8 interaction analysis. Representative composite and zoomed in images (indicated with the dotted white box) of the composite images of pre-incubation (A) or co-incubation (B) studies performed on LPS/IFN γ -activated RAW264.7 macrophages. RhB-labelled AP8ii (10 μmol/L) that was either co-incubated with R243 (10 μmol/L), CCL1 (100 ng/mL), or the anti-CCR8 antibody (5.0 μg/mL) for 4 h or R243 (10 μmol/L), CCL1 (100 ng/mL), or the anti-CCR8 antibody (5.0 μg/mL) was preincubated for 2 h followed by incubation with AP8ii (10 μmol/L) for 4 h. (C) Surface Plasmon Resonance analysis using the CellVysion SPR imaging system. The first two columns show the peptide interaction with LPS/IFN γ -activated RAW264.7 and THP-1 macrophages, while the last two columns show peptide binding following anti-CCR8 antibody blocking. Results from 6 independent experiments (performed in duplicates) are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01; One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques: Incubation, SPR Assay, Imaging, Binding Assay, Blocking Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Design, molecular characterization and therapeutic investigation of a novel CCR8 peptide antagonist that attenuates acute liver injury by inhibiting infiltration and activation of macrophages

doi: 10.1016/j.apsb.2025.02.018

Figure Lengend Snippet: AP8ii inhibits CCL1-induced migration and LPS/IFN γ -induced activation of macrophages in vitro . (A) Schematic representation of CCL1-induced migration assay; macrophages/monocytes are seeded in the Transwell insert with or without AP8ii, R243 or DMSO (10 μmol/L), and CCL1 (10 ng/mL) is added in the lower well. (B) Representative pictures of migrated nuclei (DAPI)-stained RAW macrophages. (C) Analyzed results of the migration assay performed on mouse RAW macrophages ( n = 3), BMDMs ( n = 3) and human THP1 monocytes ( n = 6). (D) Schematic showing the experimental setup for the experiment. (E) Graphs showing the gene expression analysis of Inos , Il6 , and Ccr8 performed on mouse RAW264.7 macrophages. The cells were activated with 100 ng/mL LPS + 10 ng/mL IFN γ with and without 10 μmol/L of CCR8 antagonizing peptide (AP8ii) or CCR8 antagonist (R243) with anti-CCR8 antibodies (CCR8AB) or 10 ng/mL IL4 and 10 ng/mL IL13 for 24 h followed by gene expression analysis. All results are presented as mean ± SEM from three independent experiments. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, One-way ANOVA with Bonferroni post hoc test.

Article Snippet: Immunofluorescent staining was performed using rabbit anti-mouse CCR8 (Cat. No. MAB-8324, Novus biologicals) and rat anti-mouse F4/80 (Cat. No. MCA497, BioRad) or CCR8 and rat anti-mouse CD11b (BioLegend).

Techniques: Migration, Activation Assay, In Vitro, Staining, Gene Expression

Journal: The American journal of pathology

Article Title: Chemokine (C-C Motif) Ligand 1 Derived from Tumor-Associated Macrophages Contributes to Esophageal Squamous Cell Carcinoma Progression via CCR8-Mediated Akt/Proline-Rich Akt Substrate of 40 kDa/Mammalian Target of Rapamycin Pathway.

doi: 10.1016/j.ajpath.2021.01.004

Figure Lengend Snippet: Figure 3 Chemokine (C-C motif) ligand 1 (CCL1), secreted by tumor-associated macrophage (TAM)elike macrophages, promoted TE-8, TE-9, and TE-15 cell migration and invasion. Transwell migration and invasion assays were performed to confirm the effect of neutralizing antibodies against CCL1 or CCR8 on co-culturing TE-8, TE-9, or TE-15 cells with TAM-like macrophages (TAM8, TAM9, or TAM15, respectively). A: Cells in the upper chamber were treated with a neutralizing antibody against CCR8 (anti-CCR8; 10 ng/mL) or rat IgG (10 ng/mL; control IgG; negative control); migrated cells were counted after 24 hours. B: Neutralizing antibody against CCL1 (anti-CCL1; 0.2 mg/mL) or mouse IgG (0.2 mg/mL; control IgG; negative control) was added to the lower chamber; migrated cells were counted after 24 hours. C: Cells in the upper chamber were treated with anti-CCR8 (10 ng/mL) or rat IgG (10 ng/mL; control IgG; negative control); invaded cells were counted after 48 hours. D: Anti-CCL1 (0.2 mg/mL) or mouse IgG (0.2 mg/mL; control IgG; negative control) was added to the lower chamber; invaded cells were counted after 48 hours. Results are expressed as means SEM (AeD). *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: The following antibodies were used for antigen detection in ESCC tissues: rabbit antibody against CCL1 (1:50; number HPA049861; Atlas Antibody, Stockholm, Sweden) and rabbit antibody against CCR8 (1:100; number NBP2-15768; Novus Biologicals, Littleton,

Techniques: Migration, Control, Negative Control

Journal: The American journal of pathology

Article Title: Chemokine (C-C Motif) Ligand 1 Derived from Tumor-Associated Macrophages Contributes to Esophageal Squamous Cell Carcinoma Progression via CCR8-Mediated Akt/Proline-Rich Akt Substrate of 40 kDa/Mammalian Target of Rapamycin Pathway.

doi: 10.1016/j.ajpath.2021.01.004

Figure Lengend Snippet: Figure 1 Up-regulation of chemokine (C-C motif) ligand 1 (CCL1) expression in tumor-associated macrophage (TAM)elike macrophages. A: Expression of CCL1 mRNA in peripheral blood monocyte (PBMo)ederived macrophages, TAM-like macrophages (TAM8, TAM9, and TAM15), and TE-8, TE-9, and TE-15 cells was determined using quantitative real-time PCR. Data were normalized to GAPDH levels (internal control). Assays were performed in triplicate. B: CCL1 con- centration in the supernatant of PBMo-derived macrophages, TAM-like macrophages (TAM8, TAM9, and TAM15), and TE-8, TE-9, and TE-15 cells. Protein levels were measured using an enzyme-linked immunosorbent assay. Assays were performed in triplicate. C: Expression of CCL1 in PBMo-derived macrophages and TAM-like macrophages (TAM8, TAM9, and TAM15) was confirmed by immunofluorescence using an anti-CCL1 antibody (green). Nuclei were stained with DAPI (blue). Macrophages were stained using an anti-CD204 antibody (red). Data are expressed as means SEM (A and B). *P < 0.05, **P < 0.01, and ***P < 0.001. Scale bars Z 50 mm (C).

Article Snippet: ESCC tissues were incubated with rabbit antibody against CCL1 (1:50; number HPA049861; Atlas Antibody)/CCR8 (1:100; number NBP2-15768; Novus Biologicals), mouse antibody against CD204 (1:100; number KT022; Trans Genic Inc.), mouse antibody against forkhead box P3 (1:100; number 20034; Abcam), or sheep antibody against fibroblast activation protein (1:100; number AF3715; R&D systems) at 4 C overnight.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: The American journal of pathology

Article Title: Chemokine (C-C Motif) Ligand 1 Derived from Tumor-Associated Macrophages Contributes to Esophageal Squamous Cell Carcinoma Progression via CCR8-Mediated Akt/Proline-Rich Akt Substrate of 40 kDa/Mammalian Target of Rapamycin Pathway.

doi: 10.1016/j.ajpath.2021.01.004

Figure Lengend Snippet: Figure 3 Chemokine (C-C motif) ligand 1 (CCL1), secreted by tumor-associated macrophage (TAM)elike macrophages, promoted TE-8, TE-9, and TE-15 cell migration and invasion. Transwell migration and invasion assays were performed to confirm the effect of neutralizing antibodies against CCL1 or CCR8 on co-culturing TE-8, TE-9, or TE-15 cells with TAM-like macrophages (TAM8, TAM9, or TAM15, respectively). A: Cells in the upper chamber were treated with a neutralizing antibody against CCR8 (anti-CCR8; 10 ng/mL) or rat IgG (10 ng/mL; control IgG; negative control); migrated cells were counted after 24 hours. B: Neutralizing antibody against CCL1 (anti-CCL1; 0.2 mg/mL) or mouse IgG (0.2 mg/mL; control IgG; negative control) was added to the lower chamber; migrated cells were counted after 24 hours. C: Cells in the upper chamber were treated with anti-CCR8 (10 ng/mL) or rat IgG (10 ng/mL; control IgG; negative control); invaded cells were counted after 48 hours. D: Anti-CCL1 (0.2 mg/mL) or mouse IgG (0.2 mg/mL; control IgG; negative control) was added to the lower chamber; invaded cells were counted after 48 hours. Results are expressed as means SEM (AeD). *P < 0.05, **P < 0.01, and ***P < 0.001.

Article Snippet: ESCC tissues were incubated with rabbit antibody against CCL1 (1:50; number HPA049861; Atlas Antibody)/CCR8 (1:100; number NBP2-15768; Novus Biologicals), mouse antibody against CD204 (1:100; number KT022; Trans Genic Inc.), mouse antibody against forkhead box P3 (1:100; number 20034; Abcam), or sheep antibody against fibroblast activation protein (1:100; number AF3715; R&D systems) at 4 C overnight.

Techniques: Migration, Control, Negative Control