Journal: International journal of molecular medicine

Article Title: Role of angiotensin II type 1 receptor in cerebral aneurysm formation in rats.

doi: 10.3892/ijmm_00000239

Figure Lengend Snippet: Figure 5. Immunostaining of NF-κB, MCP-1, MMP-2 and MMP-9 in cerebral aneurysmal walls and zymography of MMP-2 and MMP-9 activities. Immunostaining of DNA binding form of NF-κB p65 subunit (C and D), MCP-1 (E and F), MMP-2 (G and H) and MMP-9 (I and J) in cerebral aneurysmal walls of rats with vehicle (C, E, G, I) or valsartan treatment (D, F, H, J). (A and B) Elastica van Gieson staining (EvG) of serial section as C (A) or D (B). Bar: 30 μm. (K) Representative image of gelatine zymography from five independent experiments.

Article Snippet: Primary antibodies used in this study are listed below: rabbit polyclonal anti-monocyte chemoattractant protein (MCP-1) antibody (Santa Cruz, Santa Cruz, CA), mouse monoclonal anti-smooth muscle ·-actin antibody (Lab Vision, Fermont, CA), goat polyclonal anti-CD68 antibody (Santa Cruz), mouse monoclonal anti-NF-κB p65 subunit antibody which recognizes only DNA-binding form (Chemicon, Temecula, CA), rabbit polyclonal anti-matrix metalloproteinase (MMP)-2 antibody (Santa Cruz), goat polyclonal anti-MMP-9 antibody (Santa Cruz), rabbit polyclonal anti-AT1R antibody (Santa Cruz), and rabbit polyclonal anti-AT2R antibody (Santa Cruz).

Techniques: Immunostaining, Zymography, Binding Assay, Staining

Journal: International journal of molecular medicine

Article Title: Role of angiotensin II type 1 receptor in cerebral aneurysm formation in rats.

doi: 10.3892/ijmm_00000239

Figure Lengend Snippet: Figure 4. Effect of valsartan on MCP-1, MMP-2 and MMP-9 mRNA expression. (A-C) Result of quantitative real-time PCR analysis of MCP-1 (A), MMP-2 (B) and MMP-9 (C) with vehicle or valsartan treatment (n=6).

Article Snippet: Primary antibodies used in this study are listed below: rabbit polyclonal anti-monocyte chemoattractant protein (MCP-1) antibody (Santa Cruz, Santa Cruz, CA), mouse monoclonal anti-smooth muscle ·-actin antibody (Lab Vision, Fermont, CA), goat polyclonal anti-CD68 antibody (Santa Cruz), mouse monoclonal anti-NF-κB p65 subunit antibody which recognizes only DNA-binding form (Chemicon, Temecula, CA), rabbit polyclonal anti-matrix metalloproteinase (MMP)-2 antibody (Santa Cruz), goat polyclonal anti-MMP-9 antibody (Santa Cruz), rabbit polyclonal anti-AT1R antibody (Santa Cruz), and rabbit polyclonal anti-AT2R antibody (Santa Cruz).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Biology

Article Title: Unveiling Lovastatin’s Anti-Inflammatory Potential in Mouse’s Brain during Acute Trypanosoma cruzi Infection

doi: 10.3390/biology13050301

Figure Lengend Snippet: Antibodies used for Western blot analysis ( Figures S1–S4 in Supplementary Materials ).

Article Snippet: Primary antibody: Anti-MCP-1 , Abcam , ab702 , 1:1000 , 20 μg.

Techniques: Western Blot, Protein Concentration

Journal: Biology

Article Title: Unveiling Lovastatin’s Anti-Inflammatory Potential in Mouse’s Brain during Acute Trypanosoma cruzi Infection

doi: 10.3390/biology13050301

Figure Lengend Snippet: Analysis of inflammatory mediators by Western blot in mice’s brain at 15 dpi. Infection led to an increase in ICAM-1 levels in YNT group; lovastatin treatment prevented this increase ( A ). Neither infection nor lovastatin treatment altered VCAM-1 levels ( B ) and MCP-1 levels ( C ). NI, non-infected; Y, infected; NT, non-treated; LOV, treated with lovastatin (20 mg/kg/day); dpi, days post infection. Asterisk compared to NINT; Sharp compared to YNT; # p < 0.05, ** p < 0.01. n = 3/4 animals per experiment in 2 independent experiments.

Article Snippet: Primary antibody: Anti-MCP-1 , Abcam , ab702 , 1:1000 , 20 μg.

Techniques: Western Blot, Infection

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: CCL2 immunohistochemical staining in TCs and ICs. Upper row, ( A ) TCs, positive (ICs, negative); ( B ) Positive ICs with >6% CCL2 positivity (TCs, negative); Lower row, ( C ) TCs and ICs, negative; All photos are at 20× magnification.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Immunohistochemical staining, Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan–Meier analysis: Association of CCL2 staining in TCs with mean OS, mean DSS, or mean RFS.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan–Meier analysis: Association between CCL2 expression in TCs and prognosis. Positive CCL2 expression in TCs was associated with a shorter mean OS ( p = 0.004), mean DSS ( p = 0.036), and mean RFS ( p = 0.047) than negative CCL2 expression.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Univariate and multivariate Cox’s regression analysis: Association of CCL2 staining in TCs with mean OS, mean DSS, or mean RFS.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan–Meier analysis: Association of CCL2 staining in ICs with mean OS, mean DSS, or mean RFS.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan-Meier analysis: Association between CCL2 expression in ICs and prognosis. Positive CCL2 expression in ICs was associated with a longer mean OS (P = 0.032), mean DSS (P = 0.001) and mean RFS (P = 0.001) than negative CCL2 expression.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Univariate and multivariate Cox’s regression analysis: Association of CCL2 staining in ICs with mean OS, mean DSS, or mean RFS.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan-Meier analysis: Association of CCL2 expression stratified by tumor stage (pT2 vs. pT3+4) in ICs and prognosis. Analysis of patients in the pT2 group revealed that positive CCL2 expression was associated with longer mean DSS ( p = 0.033) and mean RFS ( p = 0.022) than negative CCL2 expression. Comparably for patients in the pT3+4 group, CCL2 expression was associated with longer mean DSS ( p = 0.030) and mean RFS ( p = 0.034).

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan-Meier analysis: Association of CCL2 expression stratified by lymph node stage; (N0 vs. N1+2) in ICs and prognosis. In the N0 group, CCL2-positive patients showed a longer mean OS ( p = 0.005), mean DSS and mean RFS (both p < 0.001) than CCL2-negative patients. However, in the N1+2 group, CCL2-positive patients had a shorter mean OS ( p = 0.001), mean DSS ( p = 0.031) and RFS ( p = 0.013).

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan-Meier analysis: Association of CCL2 expression with ICs and prognosis in the patient group not treated with chemotherapy. In the chemotherapy untreated (CT–) group, CCL2 positivity was positively associated with OS ( p = 0.012), DSS ( p < 0.001) and RFS ( p < 0.001). However, there was no association between CCL2 staining and OS, DSS or RFS in the chemotherapy-treated group (CT+).

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing, Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Multivariate Cox’s regression analysis: Interactions of the parameters CCL2 staining, lymph node status, and application of chemotherapy.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Staining

Journal: Cancers

Article Title: CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

doi: 10.3390/cancers12051253

Figure Lengend Snippet: Kaplan-Meier analysis: Association of CCL2 expression in the basal subtype in ICs and prognosis. In the basal subtype, CCL2 positive staining was associated with a better mean DSS ( p = 0.032) and mean RFS ( p = 0.044) than CCL2 negative staining. However, no association between CCL2 staining and prognosis was found in the other molecular subtypes.

Article Snippet: Briefly, after heat pretreatment at 120 °C for 5 min with TE–buffer pH 9 and peroxidase blocking (Dako, Hamburg, Germany), primary antibody against CCL2 (monoclonal mouse IgG1, clone 2D8, Cat.-No. AM06749PU-N, dilution 1:15,000; Acris Antibodies, Herford, Germany) was applied for 30 min.

Techniques: Expressing, Staining, Negative Staining

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: Intracranial injections of AAV9-RFP or AAV9-CCL2 in cortex and hippocampus in rTg4510 mice. Five-mo-old rTg4510 mice underwent intracranial bilateral injection of rAAV9-CCL2 ( n = 6) or red fluorescent protein (RFP, n = 6) in both the hippocampus and the anterior cortex. Two months after the intracranial injections, brain tissue was collected. Micrographs represent the immunohistochemistry staining for CCL2 in RFP injected (A–C) and CCL2- injected animals (D–F) . Higher magnification shows cell localization of CCL2 (insets). (G) Percent positive area of distribution in anterior cortex (ACX, boxed area), hippocampus (DG and CA3, boxed area) and entorhinal cortex (EC) of injected animals ( n = 6, ** p < 0.01 and *** p < 0.001). (H) CCL2 levels were measured in the hippocampus of injected mice by multiplex assay and the results were normalized to the amount of protein (ng/mg of protein). Student's t -test were performed between RFP and CCL2 groups for each dependent variable and each brain region separately. Scale bar represents 1,000 μm in (A,D) , 200 μm in (B,C,E,F) and 20 μm in insets.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Injection, Immunohistochemistry, Staining, Multiplex Assay

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: CCL2 overexpression does not affect microglia abundance in rTg4510. Micrograph representation of hippocampal area (A,B) and cortical area (C,D) stained for Iba-1 in RFP injected animals (A,C) and CCL2 injected animals (B,D) . Quantification of positive area stained is presented for anterior cortex (ACX), hippocampus (HPC) entorhinal cortex (EC) and meninges (MNG) for Iba-1 (mean ± S.E.M, n = 6) (E) . Student's t -test were prformed between RFP and CCL2 groups for each dependent variable and each brain region separately ( n = 6). The scale bar represents 50 μm in panels and 20 μm in insets. Meningeal area is outlined in the cortical images ( C,D , m).

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Over Expression, Staining, Injection

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: CCL2 overexpression induces microglia activation in rTg4510 brain. Micrograph representation of cortical area stained for CD45 and CD68 in RFP injected animals (A,C) and CCL2 injected animals (B,D) . Quantification of positive area stained is presented for anterior cortex (ACX), hippocampus (HPC) and entorhinal cortex (EC) for CD45 (E) and CD68 (F) (mean ± S.E.M, n = 6). Student's t -test were performed between RFP and CCL2 groups for each dependent variable and each brain region separately ( n = 6, * p < 0.05, ** p < 0.01). The scale bar represents 200 and 20 μm in panels and insets, respectively.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Over Expression, Activation Assay, Staining, Injection

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: Tmem119 is stably expressed and co-localizes with Iba-1 + microglia in rTg4510 mice. Imaging of fluorescence labeled microglia with (A,E) Tmem119 (green) and (B,F) Iba-1 (purple) in the hippocampus of RFP- and CCL2-injected rTg4510 mice. Nuclei is stained with DAPI (C,G) . Merged images of Tmem119 and Iba1 immunoreactivity in brain sections from treated mice (D,H) . N = 6, 8 section per animal. Scale bar, 100 μm (I,J,L,M) . High magnification (60x) co-localization images utilizing z-stack image intensity of highly ramified Tmem119 + cells and Iba-1 + microglia. Scale bar, 10 μm. (K,N) The intensity correlation analysis represented by the scatter plots of the fluorescence intensities of Tmem119 (Alexa Fluor 488) and Iba-1 (Alexa Fluor 647) of confocal z-stacks. The degree of co-localization is estimated by Mander's overlap coefficient.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Stable Transfection, Imaging, Fluorescence, Labeling, Injection, Staining

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: CCL2 induces astrocytic ramification in rTg4510 mice. (A) Images of fluorescence immunohistochemical stain of astrocytes (GFAP, green) in the cortices of rTg4510 mice following CCL2 and (B) RFP overexpression. Individual astrocytes were subjected to Sholl analysis (box inset). (C,D) Maximum intensity projection of each cell tiled micrographs extracting 2D images of astrocytes. Sholl-based metrics of arborization using a 50 μm radii area from the soma of each astrocyte measured the number of intersections. (E) Quantification of positive area stained for GFAP is presented for anterior cortex (ACX). (F) Sholl analysis retrieved curve-fitting and regression analysis of astrocytes within the region of interest, demonstrated induced astrocytic intersection in CCL2 overexpressing mice compared to the RFP mice. (G) Scattered plot of the number of astrocytic intersections in each group, Student t -test, Mann-Whitney unpaired parametric test, **** p < 0.0001.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Fluorescence, Immunohistochemical staining, Staining, Over Expression, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: Cytokine levels following expression of CCL2 in rTg4510 mice. The concentrations of Interleukin 1 alpha (IL-1α), Interleukin 1 beta (IL-1β), Interleukin 6 (IL-6), Interleukin 10 (IL-10), Interleukin 13 (IL-13), C-C motif ligand 11 (CCL11), Keratinocyte chemoattractant (KC), C-C motif ligand 3 (CCL3), C-C motif ligand 5 (CCL5), Vascular endothelial growth factor (VEGF) and Tumor necrosis factor alpha (TNF-α) were measured using the mouse cytokine/chemokine panel (MILLIPLEX MAP kit; Millipore, Billerica, MA, USA) in CCL2 injected mice and RFP controls. (mean ± S.E.M, n = 6). Statistical analysis was performed using multiple t -test analysis (* p < 0.05) with alpha = 0.05. and without assuming a consistent SD (df = N−2) followed by post-hoc test.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Expressing, Injection

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: CCL2 overexpression induces tau accumulation in rTg4510 mice. Images of immunostaining in the cortical area for H150 tau (A,B) , p-tau AT8 (C,D) and pSer396 (E,F) as well as Gallyas staining (aggregated tau G,H ) in mice injected with either rAAV9-RFP or rAAV9-CCL2. Quantification of positive area stained is shown in anterior cortex (ACX), hippocampus (HPC), entorhinal cortex (EC) for total tau H150 (I) , p-tau AT8 (J) , pSer396 (K) and Gallyas (L) . Scale bar represents 100 and 20 μm. Statistical analysis by Student t -test ( n = 6, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Over Expression, Immunostaining, Staining, Injection

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: CCL2 overexpression worsens tau pathology in rTg4510 mice. Western blots analyses of total tau (H150), and p-tau (AT180, PHF1, pSer396, and pSer199/202) in the soluble hippocampal brain fraction (A) and insoluble fraction (B) . Band pixel densitometry values normalized to GAPDH and control mice for soluble fraction (C) , and to controls for insoluble fraction (D) ( n = 6, * p < 0.05, ** p < 0.01, Student t -test). Overall, CCL2 overexpression resulted in increased high molecular weight tau in soluble fractions together with increased intensity of phosphorylated epitopes in insoluble fractions.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Over Expression, Western Blot, Molecular Weight

Journal: Frontiers in Immunology

Article Title: CCL2 Overexpression in the Brain Promotes Glial Activation and Accelerates Tau Pathology in a Mouse Model of Tauopathy

doi: 10.3389/fimmu.2020.00997

Figure Lengend Snippet: Amyloidosis following CCL2 overexpression in APP/PS1 mice. Micrograph representation of cortical area stained for amyloid beta (Aβ, A,B ) and Congo red (C,D) in GFP injected animals (A,C) and CCL2 injected animals (B,D) . Quantification of positive area stained is presented for anterior cortex amyloid beta (E) and Congo red (F) (Avg ± S.E.M, n = 10). Statistical analysis was performed using Student's t -test ( n = 6, * p < 0.05) followed by Fisher's PLSD multiple comparison test. Scale bar represents 200 μm and insets represent 20 μm.

Article Snippet: The following primary antibodies were used for immunohistochemistry: CCL2 (rat anti-CCL2; R&D Systems, Minneapolis, MN, USA), CD45 (rat anti-mouse; AbD Serotec, Raleigh, NC, USA), CD68 (Bio-Rad, Hercules, CA), Iba-1 (Wako, Richmond, VA) anti-tau phosphorylated at Ser396 (rabbit polyclonal, Anaspec, Fremont, CA), total tau H150 (rabbit polyclonal, SantaCruz Biotechnology), anti-tau phosphorylated at Ser202/Thr205 AT8 (Thermo scientific, Waltham, MA), anti-Aβ (prepared by Paul Gottschall, UAMS, Arkansas, USA).

Techniques: Over Expression, Staining, Injection, Comparison