Journal:

Article Title: Angiotensin II plays a pathogenic role in immune-mediated renal injury in mice

doi:

Figure Lengend Snippet: Expression of MCP-1. (a) Gene expression of MCP-1 during the first 14 days after the anti-GBM AS administration was examined by Northern blot analysis. Total RNA from mice at day 0 (control), 6 h, and days 1, 7, and 14 was separated on an agarose gel and probed for MCP-1. The blots were stripped and rehybridized with a cDNA probe for the 28S rRNA. Two representative results of each animal group were shown. (b) The gene expression of MCP-1 was estimated as described in Methods. The closed bars represent the AT1a+/+ mice, and the hatched bars represent the AT1a–/– mice. Data are mean ± SE in arbitrary units. *P < 0.05 compared with the AT1a+/+ mice. (c) Immunoperoxidase staining for MCP-1 at day 7 after anti-GBM AS administration. In the AT1a+/+ mice, MCP-1 expression was propagated in the glomeruli. In contrast, MCP-1 expression was reduced in the AT1a–/– mice. ×200. MCP-1, monocyte chemoattractant protein-1.

Article Snippet: The sections were incubated with the following antibodies: goat anti–human collagen type I cross-reactive with mouse collagen type I (Southern Biotechnology Associates, Birmingham, Alabama, USA), goat anti–mouse C3c (Nordic Immunological Laboratories, Tilburg, the Netherlands), and goat anti–mouse MCP-1 (Santa Cruz Biotechnology Inc., Santa Cruz, California, USA).

Techniques: Expressing, Northern Blot, Agarose Gel Electrophoresis, Immunoperoxidase Staining

Journal: Neuroscience

Article Title: Prenatal exposure to ethanol stimulates hypothalamic CCR2 chemokine receptor system: Possible relation to increased density of orexigenic peptide neurons and ethanol drinking in adolescent offspring

doi: 10.1016/j.neuroscience.2015.09.020

Figure Lengend Snippet: Antibodies used in single immunofluorescence histochemistry. Vendor for secondary antibodies: JacksonImmunoResearch Laboratories, Inc, PA

Article Snippet: Goat anti-CCL2 , 1/100 , Santa Cruz, CA , Cy3-Donkey anti-Goat IgG , 1/100.

Techniques: Immunofluorescence, Concentration Assay

Journal: Journal of Virology

Article Title: Viral Expression of CCL2 Is Sufficient To Induce Demyelination in RAG1 −/− Mice Infected with a Neurotropic Coronavirus

doi: 10.1128/jvi.79.11.7113-7120.2005

Figure Lengend Snippet: FIG. 1. Schematic diagram of recombinant J2.2 virus constructions. (A) Recombinant J2.2-v-1 (rJ2.2) was generated as described in Ma- terials and Methods. (B) To engineer a recombinant virus that ex- pressed CCL2, a CCL2-specific PCR product was cloned from RNA harvested from bone marrow cells and inserted into gene 4 of rJ2.2 using SbfI and MluI restriction sites. (C) As a control, a virus that encoded a truncated CCL2 protein was also generated (lys [AAG] to stop codon [TAG] at residue 52; rJ2.2.CCL2).

Article Snippet: Slides were blocked with 10% horse serum for 10 min and incubated overnight at 4°C with goat anti-mouse CCL2 antibody (1:200; R&D systems, Minneapolis, MN) and mouse anti-N MAb.

Techniques: Recombinant, Virus, Generated, Clone Assay, Control, Residue

Journal: Journal of Virology

Article Title: Viral Expression of CCL2 Is Sufficient To Induce Demyelination in RAG1 −/− Mice Infected with a Neurotropic Coronavirus

doi: 10.1128/jvi.79.11.7113-7120.2005

Figure Lengend Snippet: FIG. 2. Detection of CCL2 mRNA and protein in HeLa-MHVR cells infected with rJ2.2.CCL2. (A) CCL2 expression in cells infected with rJ2.2, rJ2.2.CCL2, or rJ2.2.CCL2 was examined at 16 h p.i. by RT-PCR and agarose gel electrophoresis. Lanes: 1, 100-bp DNA ladder; 2, rJ2.2; 3, rJ2.2.CCL2; 4, rJ2.2.CCL2. While the S gene was detected in all samples, the CCL2 gene was detected only in the cells infected with rJ2.2.CCL2 or rJ2.2.CCL2. (B) Sequence analysis showed that the CCL2 gene amplified from rJ2.2.CCL2-infected cells (lower panel) contained the introduced stop codon. (C) Expression of CCL2 protein was detected with anti-CCL2 antibody, as described in Materials and Methods. Whereas cells infected with rJ2.2, rJ2.2.CCL2, or rJ2.2.CCL2 all expressed the viral N protein, CCL2 was detected only in cells infected with rJ2.2.CCL2.

Article Snippet: Slides were blocked with 10% horse serum for 10 min and incubated overnight at 4°C with goat anti-mouse CCL2 antibody (1:200; R&D systems, Minneapolis, MN) and mouse anti-N MAb.

Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Sequencing

Journal: Journal of Virology

Article Title: Viral Expression of CCL2 Is Sufficient To Induce Demyelination in RAG1 −/− Mice Infected with a Neurotropic Coronavirus

doi: 10.1128/jvi.79.11.7113-7120.2005

Figure Lengend Snippet: FIG. 3. Functional CCL2 was detected in the supernatants of cells infected with recombinant J2.2.CCL2. HeLa-MHVR cells were in- fected with rJ2.2, rJ2.2.CCL2, or rJ2.2.CCL2 at an MOI of 0.5. Supernatants were harvested and analyzed as described in Materials and Methods. To confirm that CCL2 was responsible for the transmi- gration indicated in the figure, some samples were preincubated with anti-CCL2 antibody (100 g/ml) for 30 min. THP-1 cells (5 105

Article Snippet: Slides were blocked with 10% horse serum for 10 min and incubated overnight at 4°C with goat anti-mouse CCL2 antibody (1:200; R&D systems, Minneapolis, MN) and mouse anti-N MAb.

Techniques: Functional Assay, Infection, Recombinant

Journal: Journal of Virology

Article Title: Viral Expression of CCL2 Is Sufficient To Induce Demyelination in RAG1 −/− Mice Infected with a Neurotropic Coronavirus

doi: 10.1128/jvi.79.11.7113-7120.2005

Figure Lengend Snippet: FIG. 4. Infection with rJ2.2.CCL2 resulted in delayed mortality compared to infection with rJ2.2.CCL2. (A) Six-week-old RAG1/ mice were infected i.c. with 500 PFU of either rJ2.2.CCL2 (n 12) or rJ2.2.CCL2 (n 16). rJ2.2.CCL2-infected mice began to show clinical signs consistent with demyelination, including wobbly gait and hindlimb paresis, at 12 to 14 days p.i., and became moribund by 15 to 17 days p.i. In contrast, mice infected with rJ2.2.CCL2 displayed signs consistent with encephalitis, including hunching, ruffled fur, and lethargy, at 9 to 11 days p.i. and became moribund by 14 days p.i. The fraction surviving at each day p.i. is shown. (B) To determine whether the CCL2 gene was deleted from rJ2.2.CCL2 or rJ2.2.CCL2 after passage in mice, RNA was analyzed by RT-PCR using primers that flanked the inserted sequence. Only full-length products were detected.

Article Snippet: Slides were blocked with 10% horse serum for 10 min and incubated overnight at 4°C with goat anti-mouse CCL2 antibody (1:200; R&D systems, Minneapolis, MN) and mouse anti-N MAb.

Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Sequencing

Journal: Journal of Virology

Article Title: Viral Expression of CCL2 Is Sufficient To Induce Demyelination in RAG1 −/− Mice Infected with a Neurotropic Coronavirus

doi: 10.1128/jvi.79.11.7113-7120.2005

Figure Lengend Snippet: FIG. 5. Detection of demyelination in rJ2.2-infected RAG1/ mice. RAG1/ mice were infected i.c. with rJ2.2.CCL2 (A to C, G, and H) or rJ2.2.CCL2 (D to F). Mice were harvested at 12 to 14 days p.i., and serial longitudinal sections (8 m thick) of spinal cord were examined for demyelination (A and D), macrophage/microglia infiltration (B and E), and viral antigen (C and F) as described in Materials and Methods. Demyelination (A) and extensive macrophage/microglia infiltration into the white matter (B) were evident only in rJ2.2.CCL2-infected mice, but not in those infected with rJ2.2.CCL2 (D and E). However, viral antigen was uniformly distributed throughout spinal cords in both rJ2.2.CCL2- infected (C) and rJ2.2.CCL2-infected (F) mice, except in areas of demyelination (compare A and C). Axons were preserved (G) in areas of demyelination (A). No axonal staining was detected in the absence of anti-phosphoneurofilament antibody (H). Quantification of demyelination and the number of mice analyzed in these experiments are shown in Table 1. Scale bar, 250 m.

Article Snippet: Slides were blocked with 10% horse serum for 10 min and incubated overnight at 4°C with goat anti-mouse CCL2 antibody (1:200; R&D systems, Minneapolis, MN) and mouse anti-N MAb.

Techniques: Infection, Staining

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: A product of 4T1 cells upregulates MCP-1 production by fibroblasts. ( A ) Ten thousand 3T3 cells, 1 × 10 4 , 2 × 10 4 or 4 × 10 4 4T1 cells, or 1 × 10 4 3T3 cells with different numbers (1 × 10 4 , 2 × 10 4 , or 4 × 10 4 ) of 4T1 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by 0.5 mL fresh medium. After a 4-day incubation at 37 °C, cell-free culture supernatants were collected, and the concentration of MCP-1 was measured by ELISA. Data are presented as mean ± SD. *** p < 0.0001, n = 3. Representative of three independent experiments with similar results. ( B ) Fifty thousand 3T3 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by 0.5 mL fresh medium containing different concentrations of 4T1-sup and incubated for 24 h at 37 °C. Cell-free culture supernatants were collected, and the concentration of MCP-1 was measured by ELISA. Data are presented as mean ± SD. ** p < 0.01, **** p < 0.00001. n = 3. Representative of three independent experiments with similar results. ( C ) Fifty thousand lung fibroblasts isolated from two normal BALB/c mice were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by a fresh medium containing different concentrations of 4T1-sup. After a 24-h incubation, cell-free culture supernatants were collected, and the concentration of MCP-1 was measured by ELISA. Data are presented as mean ± SD. **** p < 0.00001, n = 3.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: Increased MCP-1 production by fibroblasts in response to 4T1-sup was due to PDGFs. ( A ) Fifty thousand 3T3 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by 0.5 mL fresh medium containing 50% ( v / v ) of 4T1-sup and incubated for 1, 3, 6 or 12 h at 37 °C. Total RNA was isolated, and the expression of Mcp-1 or Kc/Cxcl1 mRNA was evaluated by qRT-PCR. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 3. Representative of two independent experiments with similar results. ( B ) Total RNA was isolated from 4T1 cells, and the expression of Pdgfa , b , c , and d mRNA was evaluated by RT-PCR. Representative of two independent experiments with similar results. ( C ) Ten μg of 4T1 cell lysate was loaded onto a polyacrylamide gel, and SDS-PAGE was performed. Proteins were transferred onto a membrane, and the presence of PDGF-A was examined by Western blotting. Representative of three independent experiments with similar results. ( D ) Fifty thousand 3T3 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by 0.5 mL fresh medium containing 100 ng/mL recombinant mouse PDGF-AA or PDGF-BB. After a 24-h incubation at 37 °C, cell-free culture supernatants were collected, and the concentration of MCP-1 was measured by ELISA. Data are presented as mean ± SD. ** p < 0.01, **** p < 0.00001, n = 3. Representative of four independent experiments with similar results.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Incubation, Isolation, Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, SDS Page, Membrane, Western Blot, Recombinant, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: Pretreatment of 3T3 cells with crenolanib inhibited increased MCP-1 production induced by 4T1-sup. ( A ) Fifty thousand 3T3 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by a fresh medium containing DMSO or 0.2 or 2 μM crenolanib. After a 30-min incubation at 37 °C, the same volume of medium or 4T1-sup was added and incubated for an additional 24 h. Cell-free culture supernatants were collected, and the concentration of MCP-1 was measured by ELISA. Data are presented as mean ± SD. **** p < 0.00001, n = 3. Representative of three independent experiments with similar results. ( B ) Fifty thousand 3T3 cells were seeded into 24-well plates. After overnight incubation at 37 °C, the medium was replaced by a fresh medium containing DMSO or 0.2 or 2 μM crenolanib. After a 30-min incubation at 37 °C, the same volume of medium or 4T1-sup was added and incubated for an additional 24 h. Total RNA was isolated, and the expression of MCP-1 mRNA was evaluated by qRT-PCR. Data are presented as mean ± SD. * p < 0.05, n = 3. Representative of three experiments with similar results.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Isolation, Expressing, Quantitative RT-PCR

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: Treatment with trapidil or crenolanib had no effect on MCP-1 production or tumor growth. ( A ) One hundred thousand 4T1 cells were injected into a mammary pad of WT mice. Five days later, when tumors were clearly detectable, 100 μL of 10 μM crenolanib in PBS was intratumorally injected five times every 12 h. Mice were euthanized 4 h after the final injection, and the weight of the tumor (left panel) and the expression of Mcp-1 mRNA in tumors (middle panel) and serum MCP-1 concentration (right panel) were measured. Data are presented as mean ± SD. n = 3 for each group. Representative of two independent experiments with similar results. ( B ) One hundred thousand 4T1 cells were injected into a mammary pad of WT BALB/c mice. Five days later, when tumors were clearly detectable, trapidil (20 mg/kg) in 100 μL PBS was intraperitoneally injected twice a day for 7 days. Mice were euthanized on the next day, and tumor weight and serum MCP-1 concentration were measured. Data are presented as mean ± SD. n = 3 for each group. Representative of two independent experiments with similar results. ( C ) One hundred thousand A8 cells were injected into a mammary pad of WT mice. Five days later, when tumors were clearly detectable, 500 μL of 10 μM crenolanib in PBS was injected i.p. twice a day for 3 days. To directly block PDGFRs in a tumor, 100 μL of 10 μM crenolanib in PBS was injected i.t. three times every 12 h. Mice were euthanized 12 h after the final injection, and the weight of the tumor ( left panel), the expression of Mcp-1 mRNA in tumors ( middle panel) and serum MCP-1 concentration ( right panel) were measured. Data are presented as mean ± SD. n = 3 for each group. Representative of two independent experiments with similar results.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Injection, Expressing, Concentration Assay, Blocking Assay

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: Histological examination and the detection of cells expressing Mcp-1 mRNA in 5-day 4T1 tumors. One hundred thousand 4T1 cells in 100 μL PBS were injected into a mammary pad of WT female BALB/c mice. Five days after the injection, the mice were euthanized, and tumors were harvested. Consecutive paraffin sections from three individual tumors were examined by different staining and in situ hybridization. Representative photos are presented. ( A ) H&E staining, scale bar = 100 μm. ( B ) IHC with anti-Ly6G Ab, scale bar = 100 μm. ( C ) IHC with anti-F4/80 Ab, scale bar = 100 μm. ( D ) IHC with anti-αSMA, scale bar = 100 μm. ( E ) ISH. Green, Mcp-1 mRNA; Red; Act2 mRNA. Peritumoral area. Scale bar = 50 μm. ( F ) ISH. Green, Mcp-1 mRNA; Red; Act2 mRNA. Intratumoral area. Scale bar = 50 μm. ( G ) ISH. Green, Mcp-1 mRNA; Red; Adgre1 mRNA. Peritumoral area. Scale bar = 50 μm. ( H ) ISH. Green, Mcp-1 mRNA; Red; Adgre1 mRNA. Intratumoral area. Scale bar = 50 μm.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Injection, Staining, In Situ Hybridization

Journal: Current Issues in Molecular Biology

Article Title: Crosstalk between Cancer Cells and Fibroblasts for the Production of Monocyte Chemoattractant Protein-1 in the Murine 4T1 Breast Cancer

doi: 10.3390/cimb43030122

Figure Lengend Snippet: Detection of cells expressing Mcp-1 mRNA in 14-day 4T1 tumors. One hundred thousand 4T1 cells in 100 μL PBS were injected into a mammary pad of WT female BALB/c mice. Fourteen days after the injection, the mice were euthanized, and tumors were harvested. ( A ) Consecutive paraffin sections from three individual tumors were examined for the expression of Mcp-1 (green) and Act2 (upper panels) or Adgre1 (red) mRNA (lower panels) by ISH. Representative photos are presented. Scale bar = 50 μm. ( B ) One million peritoneal exudate macrophages were incubated for 24 h in the absence or presence of necrotic 4T1 cells, and the concentration of MCP-1 in the cell-free supernatants was measured by ELISA. Data are presented as mean ± SD. n = 3, ** p < 0.01. ( C ) One million peritoneal exudate macrophages were incubated for 24 h in the absence or presence of 50 or 100 μL of necrotic GM-CSF-deficient A8 cells, and the concentration of MCP-1 in the cell-free supernatants was measured by ELISA. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, n = 3. Representative of three independent experiments with similar results.

Article Snippet: The concentration of MCP-1 was measured using an ELISA kit specific for mouse MCP-1 (BioLegend) or by sandwich ELISA using anti-mouse MCP-1 antibodies (R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Injection, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Theranostics

Article Title: Visualization of Tumor-Immune Interaction - Target-Specific Imaging of S100A8/A9 Reveals Pre-Metastatic Niche Establishment

doi: 10.7150/thno.17138

Figure Lengend Snippet: Secretion of S100A8/A9 by pro-inflammatory monocytes is under control of CCL2. ( a ) S100A8/A9 concentration in supernatant of FACS-sorted CCR2 + CX3CR1 low pro-inflammatory monocytes (n=5) and Gr-1 + CD115 neg (n=4) from spleens of 67NR and 4T1.2 tumor-bearing mice stimulated 48h with recombinant mouse CCL2. S100A8/A9 concentration was determined by ELISA in two independent experiments. ( b ) Axial and coronal images from SPECT analysis and relative in vivo tracer uptake in 4T1.2 tumor-bearing mice treated with an isotype control antibody (IgG) or a blocking anti-CCL2 antibody. The frequency of CCR2 + CX3CR1 low and average relative number of CCR2 + CX3CR1 low /10 6 live cells in spleens ( c ) and lungs ( d ) (n=4, for each organ and treatment analysed in two independent experiments 14d after tumor induction) is reduced under CCL2-blockade. Mean ± SE: * p <0.05.

Article Snippet: For CCL2 blocking, 4T1.2 tumor-bearing mice received an intraperitoneal injection of 100µg goat polyclonal anti-CCL2 antibody (R&D, Abingdon, UK) every other day beginning on day 4 after tumor inoculation, following established protocols .

Techniques: Concentration Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Single Photon Emission Computed Tomography, In Vivo, Blocking Assay

Journal: Theranostics

Article Title: Visualization of Tumor-Immune Interaction - Target-Specific Imaging of S100A8/A9 Reveals Pre-Metastatic Niche Establishment

doi: 10.7150/thno.17138

Figure Lengend Snippet: S100A8/A9-induced immune remodelling in lungs of mice 4T1.2-tumor bearing mice predicts metastasis. ( a ) Exemplary axial and coronal in vivo images from SPECT examination and relative tracer accumulation graphs of healthy control animals and 67NR or 4T1.2 tumor-bearing mice (d10 after tumor induction) after injection of the S100A8/A9-specific tracer or unspecific IgG to control for perfusion effects. While the unspecific IgG does not show any differences between the three groups, S100A9-SPECT reveals ongoing monocytes activation and immune remodelling in the 4T1.2 tumor-bearing mice, reflected by a strong tracer-accumulation. ( b ) Frequency of Gr-1 + CD115 + CCR2 + CX3CR1 low monocytes in lungs of control non tumor-bearing mice, 67NR and 4T1.2 tumor-bearing mice. The bar graph shows average frequency of Gr-1 + CD115 + and the relative number of CCR2 + CX3CR1 low /10 6 live cells from 5 independent experiments. ( c ) Expression of CD115 (blue), CCR2 (magenta) and S100A8/A9 (yellow) in frozen lung sections from 4T1.2-tumor bearing mice (n=3). Extracellular S100 signal is indicated by white arrows. ( d ) Representative plots and bar graph showing the frequency of mCherry + 4T1.2 cells in the lungs of 4T1.2 tumor-bearing mice at 10 and 20 days after tumor induction (n=4, one of two experiments shown). ( e ) Correlation between S100A8/A9 activity in the lungs of 4T1.2 tumor-bearing mice at day 10 and the frequency of mCherry + 4T1.2 tumor cells at day 21 after tumor induction (n=9). Red dots indicate animals that received anti CCL2 treatment. Mean ± SE: *** p <0.005, * p <0.05.

Article Snippet: For CCL2 blocking, 4T1.2 tumor-bearing mice received an intraperitoneal injection of 100µg goat polyclonal anti-CCL2 antibody (R&D, Abingdon, UK) every other day beginning on day 4 after tumor inoculation, following established protocols .

Techniques: In Vivo, Single Photon Emission Computed Tomography, Injection, Activation Assay, Expressing, Activity Assay