Journal: Frontiers in immunology

Article Title: Transcriptomic Analysis of Rat Macrophages.

doi: 10.3389/fimmu.2020.594594

Figure Lengend Snippet: FIGURE 1 | Generation and characterisation of rat embryonic stem cell derived macrophages. (A) Schematic diagram of rat embryonic stem cell (ESC)-derived macrophage differentiation. Confluent rat ESC are shown at day 0 (Clone DA5.2), embryoid bodies at day 7, and ESC-derived macrophages (ESDM) at day 25. Bars = 50, 100, and 50 µm, respectively. Images are representative of three repeat experiments, two replicates. (B) ESDM were cultured with or without (control) fluorescein labelled Zymosan A BioParticles. Images are representative of three repeat experiments, two replicates. Blue = nuclear DAPI staining. Bars = 10 µm. (C) Flow cytometry of permeabilized cells was used to assess purity of ESDM via CD68 expression. Quadrants were set using an isotype control. Dot plot is representative of three repeat experiments, two replicates.

Article Snippet: Cells were permeabilised using Leucoperm (AbD Serotec) according to instructions and incubated with mouse anti-rat CD68 (AbD Serotec, clone ED1, 1:200) followed by goat antimouse IgG APC (BioLegend, 1:400) and analysed on a FACSCalibur (BD).

Techniques: Derivative Assay, Cell Culture, Control, Staining, Flow Cytometry, Expressing

Journal: Frontiers in immunology

Article Title: Transcriptomic Analysis of Rat Macrophages.

doi: 10.3389/fimmu.2020.594594

Figure Lengend Snippet: FIGURE 2 | Adult rat macrophage populations. (A) Macrophages were isolated from adult male Dark Agouti rats. Images are representative of cells isolated from 10 rats. Bars = 20 µm (AM) or 50 µm (BMDM, MDM, and PM). (B) Permeabilized macrophages were analysed by flow cytometry to assess purity via CD68 expression. Quadrants were set with isotype controls. Dot plots are representative of cells isolated from three rats. (C) Alveolar macrophages were cultured with or without (control) fluorescein labelled Zymosan A BioParticles and imaged with a Zeiss AxioVert. Images are representative of cells isolated from three rats. Bar = 50 µm. (D) Macrophages were cultured with or without (control) fluorescein labelled Zymosan A BioParticles and imaged with a Zeiss LSM 710 confocal. Images are representative of cells isolated from three rats. Bar = 10 µm. AM = alveolar macrophages, BMDM = bone marrow derived macrophages, MDM = monocyte derived macrophages, PM, peritoneal macrophages.

Article Snippet: Cells were permeabilised using Leucoperm (AbD Serotec) according to instructions and incubated with mouse anti-rat CD68 (AbD Serotec, clone ED1, 1:200) followed by goat antimouse IgG APC (BioLegend, 1:400) and analysed on a FACSCalibur (BD).

Techniques: Isolation, Cytometry, Expressing, Cell Culture, Control, Derivative Assay

Journal: Cells

Article Title: Retinal Ganglion Cell Survival and Axon Regeneration after Optic Nerve Transection is Driven by Cellular Intravitreal Sciatic Nerve Grafts

doi: 10.3390/cells9061335

Figure Lengend Snippet: List of antibodies used in this study.

Article Snippet: Mouse anti-cluster differentiation 68 (ED1) , Rat macrophages , 1:200 , Serotec, Kidlington, UK.

Techniques: Plasmid Preparation

Journal: Cells

Article Title: Retinal Ganglion Cell Survival and Axon Regeneration after Optic Nerve Transection is Driven by Cellular Intravitreal Sciatic Nerve Grafts

doi: 10.3390/cells9061335

Figure Lengend Snippet: Macrophages accumulate in the retina in eyes containing ivit CSN implants at 21 days after grafting. ED1-HRP + macrophages in the vitreous body and retina in: ( A ) CON/intact, ( B ) CON/ ivit S /ONT, ( C ) ivit ASN /ONT, ( D ) ivit S /ONT ASN , ( E ) ivit S /ONT CSN , ( F ) ivit CSN /ONT, ( G ) ivit CSN /ONT ASN , ( H ) ivit ASN /ONT CSN , and ( I ) ivit CSN /ONT CSN groups. ED1 + macrophages were present in both ( J ) ivit CSN , and ( K ) ivit ASN implants; ED1 + macrophages (red) at the anastomosis site of ( L ) ONT CSN ; ( M ) ivit CSN /ONT ASN ; ( N ) ivit CSN /ONT CSN groups (scale bars in A – K = 50 μm; in L – N = 100 μm). n = 16 retinae/group.

Article Snippet: Mouse anti-cluster differentiation 68 (ED1) , Rat macrophages , 1:200 , Serotec, Kidlington, UK.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Small-diameter bacterial cellulose-based vascular grafts for coronary artery bypass grafting in a pig model

doi: 10.3389/fcvm.2022.881557

Figure Lengend Snippet: Inner layer of the vascular grafts 4 weeks after implantation. Representative images of the vascular grafts at the central part (A–C) and at the anastomosis site (D–F) stained with H&E (A,D) and immunofluorescence antibodies specific for Ve-Cadherin (red), α-SMA (cyan), laminin (green) (B,E) , and for CD31 (red) and CD68 (green) (C,F) . Nuclei were stained with DAPI (blue) (B,C,E,F) . Scale bar = 100 μm.

Article Snippet: For immunofluorescence staining, the following antibodies were used: rabbit anti-laminin (ab11575, dilution at 1:200) and goat anti-smooth muscle actin (SMA) (ab7817, dilution at 1:400) from Abcam (UK), goat anti-vascular endothelial (VE) cadherin (sc6458, dilution at 1:100) from Santa Cruz Biotechnology (Dallas, US), mouse anti-CD31 (MCA1746GA, dilution at 1:100) from AbD Serotec (Germany), mouse anti-CD68 (BA4D5, dilution at 1:50) from AbD Serotec (Germany).

Techniques: Staining, Immunofluorescence

Journal: Journal of Neurotrauma

Article Title: Sulfonylurea Receptor 1, Transient Receptor Potential Cation Channel Subfamily M Member 4, and KIR6.2:Role in Hemorrhagic Progression of Contusion

doi: 10.1089/neu.2018.5986

Figure Lengend Snippet: (A) Immunolabeling for sulfonylurea receptor 1 (SUR1) showed sparse immunoreactivity in the control specimen (CTR) vs. widespread expression in elongated structures and small round cells in a glial fibrillary acidic protein (GFAP)–negative specimen from contusion–traumatic brain injury (TBI). (B, C) Double immunolabeling for collagen IV (COLIV) (red) and SUR1 (B) or transient receptor potential cation channel subfamily M member 4 (TRPM4) (C) showed expression of SUR1 and TRPM4 in microvessels; merged images are also shown. (D-F) Double immunolabeling for CD68 (red) and SUR1 (D) or TRPM4 (E) or KIR6.2 (F) showed expression of SUR1, TRPM4 and KIR6.2 in microglia/macrophages; merged images are also shown. (G) Double immunolabeling for SUR1 (green) and TRPM4 (red) showed co-localization in a microvessel. (H) ImmunoFRET for SUR1 (red) and TRPM4 (magenta) shows co-assembly of SUR1-TRPM4 heteromers (yellow pseudocolor) in a microvessel. The findings illustrated are from the GFAP-negative specimen shown in , left, and are representative of all GFAP-negative, SUR1-positive specimens from four cases of human contusion-TBI; (four cases with GFAP-negative specimens showed no immunoreactivity for SUR1). Case #2, 11 days post-TBI; case #3, 7 days post-TBI.

Article Snippet: After labeling for one of the channel subunits, double immunolabeling was performed using either: CY3-conjugated, monoclonal anti-GFAP (1:500; C9205; Sigma-Aldrich), or monoclonal anti-RECA (MA1-81510; 1:100; HIS52; Invitrogen, ThermoFisher Scientific, Waltham, MA), or mouse anti-CD68 (ED1; 1:100; MAB1435; Millipore Sigma), to analyze cell specific expression.

Techniques: Immunolabeling, Expressing

Journal: Journal of Neurotrauma

Article Title: Sulfonylurea Receptor 1, Transient Receptor Potential Cation Channel Subfamily M Member 4, and KIR6.2:Role in Hemorrhagic Progression of Contusion

doi: 10.1089/neu.2018.5986

Figure Lengend Snippet: Glial fibrillary acidic protein (GFAP)–negative specimens from rat contusion–traumatic brain injury (TBI) exhibit sulfonylurea receptor 1 (SUR1) expression in microvessels and microglia/macrophages. (A-C) Double immunolabeling for rat endothelial cell antigen (RECA) (green) and SUR1 (A) or transient receptor potential cation channel subfamily M member 4 (TRPM4) (B) or KIR6.2 (C) at 24 h post-TBI showed expression of SUR1 and TRPM4 in microvessels; merged images are shown in the right panels (yellow). (D) Double immunolabeling for SUR1 (green) and TRPM4 (red) at 24 h post-TBI shows co-localization in microvessels. (E-G) Double immunolabeling for CD68 (green) and SUR1 (E) or TRPM4 (F) or KIR6.2 (G) at 72 h post-TBI showed expression of SUR1, TRPM4, and KIR6.2 in microglia/macrophages; merged images are also shown. The findings illustrated are representative of five specimens from rat contusion-TBI with GFAP-negative core.

Article Snippet: After labeling for one of the channel subunits, double immunolabeling was performed using either: CY3-conjugated, monoclonal anti-GFAP (1:500; C9205; Sigma-Aldrich), or monoclonal anti-RECA (MA1-81510; 1:100; HIS52; Invitrogen, ThermoFisher Scientific, Waltham, MA), or mouse anti-CD68 (ED1; 1:100; MAB1435; Millipore Sigma), to analyze cell specific expression.

Techniques: Expressing, Immunolabeling

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: CD1b-restricted GEM T cell responses are modulated by Mycobacterium tuberculosis mycolic acid meromycolate chains

doi: 10.1073/pnas.1708252114

Figure Lengend Snippet: CD1b expression within human TB granulomas. Human lung biopsies from patients with confirmed TB were stained for the macrophage marker CD68 (A) and CD1b (B). (C) Negative control with secondary antibody (Ab) and avidin biotin–peroxidase complex (ABC) detection only. (A–C, Insets) Large box is a 2.1× magnified version of the small box. (Scale bars: A–C, 50 μm.)

Article Snippet: Nonspecific staining was blocked, and primary antibodies were applied overnight at 4 °C (1:50 anti-CD1b mouse monoclonal SN13, K5 1B8, Abcam; 1:200 CD68 mouse monoclonal ED1, LifeSpan Biosciences).

Techniques: Expressing, Staining, Marker, Negative Control, Avidin-Biotin Assay