Review

flow n a trem2 apc 237920 rat igg2b r d systems fab17291a (R&D Systems)

R&D Systems is a verified supplier

R&D Systems manufactures this product

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

R&D Systems flow n a trem2 apc 237920 rat igg2b r d systems fab17291a
Flow N A Trem2 Apc 237920 Rat Igg2b R D Systems Fab17291a, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/trem2+apc+rat+anti+mouse/Human%2FMouse+TREM2+APC-conjugated+Antibody/pmc11757139__Presentation1-62-93-99
Average 95 stars, based on 66 article reviews

flow n a trem2 apc 237920 rat igg2b r d systems fab17291a - by Bioz Stars, 2026-09

95/100 stars

Images

Expressing:

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis
Article Snippet: Data were processed in FlowJo (Tree Star) or Cytek SpectroFlo software.Data were processed in FlowJo (Tree Star) or Cytek SpectroFlo software.. The following antibodies were used: Trem2 APC rat anti-mouse (clone 237920, R&D Systems); Trem2 FITC rat anti-mouse (clone 78.18, eBioscience); CD68 rat anti-mouse (clone FA-11, BioLegend); Ki67 rabbit anti-mouse (clone SP6, Abcam); CD45 BV480 rat anti-mouse (clone 30-F11, BioLegend); CD11b BV605 rat anti-mouse (clone M1/70, BioLegend); Ly6G BV785 rat anti-mouse (clone 1A8, BioLegend); Ly6C BV421 rat anti-mouse (clone HK1.4, BioLegend); CD115 PerCPCy5.5 rat anti-mouse (clone AFS98, BioLegend); TCRβ APC hamster anti-mouse (clone H57–597, BioLegend); CD19 FITC rat anti-mouse (clone 1D3, BioLegend); and sXBP1 AF647 rat anti-mouse (clone E9V3E, Cell Signaling Technology).

CRISPR:

Knock-Out:

Incubation:

Isolation:

Flow Cytometry:

Staining:

Cell Culture:

Control:

Immunohistochemical staining:

Irradiation:

Derivative Assay:

Labeling:

Injection:

Multiplex Assay:

Immunofluorescence:

TUNEL Assay:

RNA Sequencing:

Lactate Dehydrogenase Assay:

Two Tailed Test:

Positive Control:

Fluorescence:

Activation Assay:

Image Search Results

a ) Clustering of all cell subsets from human atherosclerotic endarterectomy samples (Fernandez et. al., Nat Med 2019). b ) Clustering of monocyte/macrophage populations from human atherosclerotic endarterectomy samples (Fernandez et. al., Nat Med 2019). c ) PTPRC and CD14 expression of clustered monocyte/macrophage populations from 3b. d ) Foamy macrophage gene (FABP5, LGALS3) expression of clustered monocyte/macrophage populations from 3b. e ) Inflammatory macrophage gene (IL1B, NLRP3) expression of clustered monocyte/macrophage populations from 3b. f ) TREM2 expression of clustered monocyte/macrophage populations from 3b. g ) Volcano plot of enrichment of genes from samples from either asymptomatic or symptomatic patients. TREM2 in red. DEGs were determined by Wald test with DESeq2.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) Clustering of all cell subsets from human atherosclerotic endarterectomy samples (Fernandez et. al., Nat Med 2019). b ) Clustering of monocyte/macrophage populations from human atherosclerotic endarterectomy samples (Fernandez et. al., Nat Med 2019). c ) PTPRC and CD14 expression of clustered monocyte/macrophage populations from 3b. d ) Foamy macrophage gene (FABP5, LGALS3) expression of clustered monocyte/macrophage populations from 3b. e ) Inflammatory macrophage gene (IL1B, NLRP3) expression of clustered monocyte/macrophage populations from 3b. f ) TREM2 expression of clustered monocyte/macrophage populations from 3b. g ) Volcano plot of enrichment of genes from samples from either asymptomatic or symptomatic patients. TREM2 in red. DEGs were determined by Wald test with DESeq2.

Article Snippet: The following antibodies were used: Trem2 APC rat anti-mouse (clone 237920, R&D Systems); Trem2 FITC rat anti-mouse (clone 78.18, eBioscience); CD68 rat anti-mouse (clone FA-11, BioLegend); Ki67 rabbit anti-mouse (clone SP6, Abcam); CD45 BV480 rat anti-mouse (clone 30-F11, BioLegend); CD11b BV605 rat anti-mouse (clone M1/70, BioLegend); Ly6G BV785 rat anti-mouse (clone 1A8, BioLegend); Ly6C BV421 rat anti-mouse (clone HK1.4, BioLegend); CD115 PerCPCy5.5 rat anti-mouse (clone AFS98, BioLegend); TCRβ APC hamster anti-mouse (clone H57–597, BioLegend); CD19 FITC rat anti-mouse (clone 1D3, BioLegend); and sXBP1 AF647 rat anti-mouse (clone E9V3E, Cell Signaling Technology).

Techniques: Expressing

a , Schematic for CRISPR knockout screening approach for oxLDL uptake. BV2 macrophages were loaded with CRISPR pooled guide library (Gouda). Cells were made foamy by overnight treatment with soluble cholesterol and then challenged for 4 h with DiI-oxLDL and sorted for DiI high and DiI low cells. Guides were sequenced from sorted populations. b , Confocal micrograph showing BV2 DiI uptake after 4-h incubation with DiI-oxLDL. Representative of two independent experiments. c , CRISPR guide enrichment comparing log-normalized enrichment in DiI high ( x axis) versus DiI low ( y axis). Gray error bands delineate guides with log fold change < 1. d , Selected gene enrichments comparing DiI low versus DiI high . e , Peritoneal macrophages were isolated from WT or Trem2 −/− mice and treated with soluble cholesterol to induce foamy cell formation. After overnight culture, cells were analyzed for Trem2 expression by flow cytometry ( n = 5 biologically independent replicates per group). Data are mean ± s.e.m. Student’s t -test, **** P < 0.0001. f , Bodipy staining for total neutral lipid accumulation was performed by flow cytometry on peritoneal macrophages from WT or Trem2 −/− mice, cultured overnight in media alone or in media with soluble cholesterol ( n = 6 biologically independent replicates for untreated and n = 4 biologically independent replicates for foamy). Data are mean ± s.e.m. Student’s t -test. g , Peritoneal macrophages were isolated from WT or Trem2 −/− mice and treated with soluble cholesterol to induce foamy cell formation. After overnight culture, cells were treated with DiI-oxLDL for 4 h and assessed for uptake by flow cytometry ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. h , CD36 expression from peritoneal macrophages isolated from WT or Trem2 −/− mice and treated with soluble cholesterol overnight to induce foamy cell formation ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. NS, not significant.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , Schematic for CRISPR knockout screening approach for oxLDL uptake. BV2 macrophages were loaded with CRISPR pooled guide library (Gouda). Cells were made foamy by overnight treatment with soluble cholesterol and then challenged for 4 h with DiI-oxLDL and sorted for DiI high and DiI low cells. Guides were sequenced from sorted populations. b , Confocal micrograph showing BV2 DiI uptake after 4-h incubation with DiI-oxLDL. Representative of two independent experiments. c , CRISPR guide enrichment comparing log-normalized enrichment in DiI high ( x axis) versus DiI low ( y axis). Gray error bands delineate guides with log fold change < 1. d , Selected gene enrichments comparing DiI low versus DiI high . e , Peritoneal macrophages were isolated from WT or Trem2 −/− mice and treated with soluble cholesterol to induce foamy cell formation. After overnight culture, cells were analyzed for Trem2 expression by flow cytometry ( n = 5 biologically independent replicates per group). Data are mean ± s.e.m. Student’s t -test, **** P < 0.0001. f , Bodipy staining for total neutral lipid accumulation was performed by flow cytometry on peritoneal macrophages from WT or Trem2 −/− mice, cultured overnight in media alone or in media with soluble cholesterol ( n = 6 biologically independent replicates for untreated and n = 4 biologically independent replicates for foamy). Data are mean ± s.e.m. Student’s t -test. g , Peritoneal macrophages were isolated from WT or Trem2 −/− mice and treated with soluble cholesterol to induce foamy cell formation. After overnight culture, cells were treated with DiI-oxLDL for 4 h and assessed for uptake by flow cytometry ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. h , CD36 expression from peritoneal macrophages isolated from WT or Trem2 −/− mice and treated with soluble cholesterol overnight to induce foamy cell formation ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. NS, not significant.

Techniques: CRISPR, Knock-Out, Incubation, Isolation, Expressing, Flow Cytometry, Staining, Cell Culture

a ) Time course analysis of DiI-oxLDL uptake in WT BV2 cells differentiated in media with 20 μg/mL of soluble cholesterol overnight prior to addition of DiI-oxLDL (n = 5 biological replicates/group). Data are mean ± S.E.M. b ) CRISPR guide enrichment by rank-order was plotted against P-value for DiI-oxLDL-low compared against DiI-oxLDL-high to identify top enriched guides. Trem2 in red. P-values calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. c ) Two sided P-value vs count and p value vs false discovery rate (FDR) for DiI-oxLDL-low compared against DiI-oxLDL-high to identify top enriched guides. p-values and FDR calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. d ) Top 15 ‘importance index’ genes associated with foamy cell commitment by Trade-seq analysis , were compared for gene rank and enrichment in CRISPR screen. Trem2 highlighted in gray. P-values calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. e) CD36 expression (left) and percent CD36 high (right) of foamy peritoneal macrophages cultured with soluble cholesterol overnight from WT (blue) and Trem2−/− mice (red). Gated on F4/80+ CD11b+ live cells (n = 5 biological replicates/group). Data are mean ± S.E.M. Student’s t-test, P = ** < 0.01. f ) SR-AI expression (left), MFI (middle) and percent SR-AI positive (right) of foamy peritoneal macrophages cultured with soluble cholesterol overnight from WT (blue) and Trem2−/− mice (red). Gated on F4/80+ CD11b+ live cells (n = 5 biological replicates/group). Data are mean ± S.E.M. Student’s t-test

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) Time course analysis of DiI-oxLDL uptake in WT BV2 cells differentiated in media with 20 μg/mL of soluble cholesterol overnight prior to addition of DiI-oxLDL (n = 5 biological replicates/group). Data are mean ± S.E.M. b ) CRISPR guide enrichment by rank-order was plotted against P-value for DiI-oxLDL-low compared against DiI-oxLDL-high to identify top enriched guides. Trem2 in red. P-values calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. c ) Two sided P-value vs count and p value vs false discovery rate (FDR) for DiI-oxLDL-low compared against DiI-oxLDL-high to identify top enriched guides. p-values and FDR calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. d ) Top 15 ‘importance index’ genes associated with foamy cell commitment by Trade-seq analysis , were compared for gene rank and enrichment in CRISPR screen. Trem2 highlighted in gray. P-values calculated using the negative-binomial model from MAGeCK package and adjusted using Benjamini-Hochberg procedure. e) CD36 expression (left) and percent CD36 high (right) of foamy peritoneal macrophages cultured with soluble cholesterol overnight from WT (blue) and Trem2−/− mice (red). Gated on F4/80+ CD11b+ live cells (n = 5 biological replicates/group). Data are mean ± S.E.M. Student’s t-test, P = ** < 0.01. f ) SR-AI expression (left), MFI (middle) and percent SR-AI positive (right) of foamy peritoneal macrophages cultured with soluble cholesterol overnight from WT (blue) and Trem2−/− mice (red). Gated on F4/80+ CD11b+ live cells (n = 5 biological replicates/group). Data are mean ± S.E.M. Student’s t-test

Techniques: CRISPR, Expressing, Cell Culture

a ) Cranial artery plaques were stained either for CD68 and Trem2 to identify co−expressing foamy macrophages within human plaques (top) or with CD68 and isotype control with secondary antibody (bottom). Representative image from 3 independent samples. b ) Carotid artery endarterectomy samples from three patients stained for isotype control or Trem2 using DAB (3,3′-Diaminobenzidine) immunohistochemical staining. Representative images from 6 independent samples.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) Cranial artery plaques were stained either for CD68 and Trem2 to identify co−expressing foamy macrophages within human plaques (top) or with CD68 and isotype control with secondary antibody (bottom). Representative image from 3 independent samples. b ) Carotid artery endarterectomy samples from three patients stained for isotype control or Trem2 using DAB (3,3′-Diaminobenzidine) immunohistochemical staining. Representative images from 6 independent samples.

Techniques: Staining, Expressing, Control, Immunohistochemical staining

a , Schematic for mixed bone marrow chimera experiment. Ldlr −/− mice were lethally irradiated and rescued by donor bone marrow from (50%) LysM cre R26 tdTomato (WT) and (50%) Trem2 −/− mice. Recipient mice were rested for 8 weeks and then fed an HFD for an additional 8 weeks to induce atherosclerosis. b , Flow cytometry gating of blood immune cells (CD45 + ) after 8-week HFD feeding, showing ratio of monocytes derived from WT (tdTomato + ) and Trem2 −/− progenitors. c , Confocal micrograph of whole-mount aorta showing tdTomato labeling (red) and CD45 (white) staining to define cellular contributions to foamy macrophages. Representative image from two independent experiments. d , Quantification of tdTomato + cells in blood compared to foamy macrophages from whole-mount aorta images ( n = 3 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. e , Foamy FACS was performed on CD64 + CD11b + macrophages isolated from mixed bone marrow chimera aorta. Macrophages were separated into tdTomato + and tdTomato − populations and then assessed for foamy representation by SSC and Bodipy (neutral lipid) staining. f , Flow cytometric overlap between tdTomato + (red) and Trem2 −/− (blue) derived macrophages from digested atherosclerotic aorta. g , Quantification derived from flow cytometric foamy FACS comparing relative contribution to foamy macrophages ( n = 4 mice per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. KO, knockout. MACS, macrophage.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , Schematic for mixed bone marrow chimera experiment. Ldlr −/− mice were lethally irradiated and rescued by donor bone marrow from (50%) LysM cre R26 tdTomato (WT) and (50%) Trem2 −/− mice. Recipient mice were rested for 8 weeks and then fed an HFD for an additional 8 weeks to induce atherosclerosis. b , Flow cytometry gating of blood immune cells (CD45 + ) after 8-week HFD feeding, showing ratio of monocytes derived from WT (tdTomato + ) and Trem2 −/− progenitors. c , Confocal micrograph of whole-mount aorta showing tdTomato labeling (red) and CD45 (white) staining to define cellular contributions to foamy macrophages. Representative image from two independent experiments. d , Quantification of tdTomato + cells in blood compared to foamy macrophages from whole-mount aorta images ( n = 3 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. e , Foamy FACS was performed on CD64 + CD11b + macrophages isolated from mixed bone marrow chimera aorta. Macrophages were separated into tdTomato + and tdTomato − populations and then assessed for foamy representation by SSC and Bodipy (neutral lipid) staining. f , Flow cytometric overlap between tdTomato + (red) and Trem2 −/− (blue) derived macrophages from digested atherosclerotic aorta. g , Quantification derived from flow cytometric foamy FACS comparing relative contribution to foamy macrophages ( n = 4 mice per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. KO, knockout. MACS, macrophage.

Techniques: Irradiation, Flow Cytometry, Derivative Assay, Labeling, Staining, Isolation, Knock-Out

a , CX3CR1 creER Trem2 flox/flox Ldlr −/− (Trem2 ΔMФ ) or littermate control mice (which included Cre − animals CX3CR1 +/+ Trem2 fl/fl Ldlr −/− and Cre + animals CX3CR1 creER/+ Trem2 fl/+ Ldlr −/− ) were fed TAM-HFD for 8 weeks ( b – e ) or 16 weeks ( f – i ). b , After 8 weeks of TAM-HFD, aortas were analyzed by en face analysis for percentage Oil Red O (ORO) staining on the arch ( n = 17 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. c , Aortic sinus plaque area measured after ORO staining in 8-week TAM-HFD samples (n = 17 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. d , Serum cholesterol levels from 8-week TAM-HFD-fed mice ( n = 11 mice per group for Cntl and n = 8 for Trem2 ΔMФ ). Data are mean ± s.e.m. e , Weight data from 8-week TAM-HFD-fed mice ( n = 9 mice pergroup). Data are mean ± s.e.m. f , En face ORO staining of aorta after 16-week TAM-HFD feeding ( n = 12 mice per group). Data are mean ± s.e.m. Student’s t -test, **** P < 0.0001. g , Aortic sinus plaque area after 16-week TAM-HFD feeding ( n = 11 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, *** P < 0.001. h , Serum cholesterol after 16-week TAM-HFD feeding ( n = 10 mice per group). Data are mean ± s.e.m. i , Weight of mice after 16-week TAM-HFD feeding ( n = 10 mice per group for Cntl and n = 7 for Trem2 ΔMФ ). Data are mean ± s.e.m.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , CX3CR1 creER Trem2 flox/flox Ldlr −/− (Trem2 ΔMФ ) or littermate control mice (which included Cre − animals CX3CR1 +/+ Trem2 fl/fl Ldlr −/− and Cre + animals CX3CR1 creER/+ Trem2 fl/+ Ldlr −/− ) were fed TAM-HFD for 8 weeks ( b – e ) or 16 weeks ( f – i ). b , After 8 weeks of TAM-HFD, aortas were analyzed by en face analysis for percentage Oil Red O (ORO) staining on the arch ( n = 17 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. c , Aortic sinus plaque area measured after ORO staining in 8-week TAM-HFD samples (n = 17 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. d , Serum cholesterol levels from 8-week TAM-HFD-fed mice ( n = 11 mice per group for Cntl and n = 8 for Trem2 ΔMФ ). Data are mean ± s.e.m. e , Weight data from 8-week TAM-HFD-fed mice ( n = 9 mice pergroup). Data are mean ± s.e.m. f , En face ORO staining of aorta after 16-week TAM-HFD feeding ( n = 12 mice per group). Data are mean ± s.e.m. Student’s t -test, **** P < 0.0001. g , Aortic sinus plaque area after 16-week TAM-HFD feeding ( n = 11 mice per group for Cntl and n = 12 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, *** P < 0.001. h , Serum cholesterol after 16-week TAM-HFD feeding ( n = 10 mice per group). Data are mean ± s.e.m. i , Weight of mice after 16-week TAM-HFD feeding ( n = 10 mice per group for Cntl and n = 7 for Trem2 ΔMФ ). Data are mean ± s.e.m.

Techniques: Control, Staining

a ) Trem2 expression and quantification from atherosclerotic aortae (n = 2 mice/group for Cntl and n = 3 for Trem2ΔMФ). Briefly, Cntl or Trem2ΔMФ mice were fed TAM-HFD for 16 weeks then aorta were harvested, digested and flow cytometry was run. Histogram was gated on live, CD45 + CD11b + CD64+ cells. b ) Flow cytometric gating strategy for identifying major blood immune cell populations. c ) Blood immune cell profiling by flow cytometry in indicated mice after 16 weeks TAM-HFD feeding (n = 12 mice/group for Cntl and n = 6 for Trem2ΔMФ). Data are mean ± S.E.M. Student’s t-test. d ) Classical monocyte bead uptake in the blood was measured by flow cytometry 24 hours after i.v. bead injection in indicated strains after 16 weeks TAM-HFD feeding (n = 7 mice/group for Cntl and n = 5 for Trem2ΔMФ). Data are mean ± S.E.M. Student’s t-test.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) Trem2 expression and quantification from atherosclerotic aortae (n = 2 mice/group for Cntl and n = 3 for Trem2ΔMФ). Briefly, Cntl or Trem2ΔMФ mice were fed TAM-HFD for 16 weeks then aorta were harvested, digested and flow cytometry was run. Histogram was gated on live, CD45 + CD11b + CD64+ cells. b ) Flow cytometric gating strategy for identifying major blood immune cell populations. c ) Blood immune cell profiling by flow cytometry in indicated mice after 16 weeks TAM-HFD feeding (n = 12 mice/group for Cntl and n = 6 for Trem2ΔMФ). Data are mean ± S.E.M. Student’s t-test. d ) Classical monocyte bead uptake in the blood was measured by flow cytometry 24 hours after i.v. bead injection in indicated strains after 16 weeks TAM-HFD feeding (n = 7 mice/group for Cntl and n = 5 for Trem2ΔMФ). Data are mean ± S.E.M. Student’s t-test.

Techniques: Expressing, Flow Cytometry, Injection

a , Following the schematic in , CX3CR1 creER Trem2 flox/flox Ldlr −/− (Trem2 ΔMФ ) or littermate control mice were treated continuously with TAM-HFD for the indicated times. b , Serum from 8-week TAM-HFD-fed mice were assessed for cytokine levels by multiplex assay ( n = 10 mice per group). Data are mean ± s.e.m. c , Serum from 16-week TAM-HFD-fed mice were assessed for cytokine levels by multiplex assay ( n = 9 mice per group). Data are mean ± s.e.m. d , Blood immune cells were assessed after 16 weeks of TAM-HFD by flow cytometry ( n = 12 mice per group for Cntl and n = 6 for Trem2 ΔMФ ). Data are mean ± s.e.m. e , Monocyte recruitment was assessed by bead labeling and recruitment experiment—images from representative histologic and immunofluorescence images with lipid content (red) and beads (green). Representative image from two independent experiments. f , Quantification of plaque-associated beads that were counted per section for 8-week or 16-week TAM-HFD experiments from experiments in ( n = 7 mice per group for Cntl and n = 5 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test. NS, not significant; ORO, Oil Red O.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , Following the schematic in , CX3CR1 creER Trem2 flox/flox Ldlr −/− (Trem2 ΔMФ ) or littermate control mice were treated continuously with TAM-HFD for the indicated times. b , Serum from 8-week TAM-HFD-fed mice were assessed for cytokine levels by multiplex assay ( n = 10 mice per group). Data are mean ± s.e.m. c , Serum from 16-week TAM-HFD-fed mice were assessed for cytokine levels by multiplex assay ( n = 9 mice per group). Data are mean ± s.e.m. d , Blood immune cells were assessed after 16 weeks of TAM-HFD by flow cytometry ( n = 12 mice per group for Cntl and n = 6 for Trem2 ΔMФ ). Data are mean ± s.e.m. e , Monocyte recruitment was assessed by bead labeling and recruitment experiment—images from representative histologic and immunofluorescence images with lipid content (red) and beads (green). Representative image from two independent experiments. f , Quantification of plaque-associated beads that were counted per section for 8-week or 16-week TAM-HFD experiments from experiments in ( n = 7 mice per group for Cntl and n = 5 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test. NS, not significant; ORO, Oil Red O.

Techniques: Control, Multiplex Assay, Flow Cytometry, Labeling, Immunofluorescence

a , Confocal micrograph showing CD68 staining (green) and DAPI (blue) for macrophage area in Cntl or Trem2-deficent mice after 16-week TAM-HFD feeding. Representative image from two independent experiments. b , Quantification of CD68 + macrophage area per section in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01. c , Quantification of the percentage of plaque that is macrophages (CD68 + ) in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test. d , Confocal micrograph showing Ki67 staining (magenta) and CD68 staining (green) for proliferation in Cntl or Trem2-deficient mice after 16-week TAM-HFD feeding. Representative image from two independent experiments. e , Quantification of Ki67 + macrophages (CD68 + ) per section in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group for 8-week TAM-HFD, n = 7 mice per group for Cntl 16-week TAM-HFD and n = 6 mice per group for 16-week TAM-HFD Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01. f , Confocal micrograph of TUNEL staining (magenta) and CD68 staining (green) for detection of dying cells within atherosclerotic lesions after 16-week TAM-HFD feeding. Representative image from two independent experiments. g , Quantification of TUNEL + macrophages (CD68 + ) per section in 8-week or 16-week TAM-HFD samples ( n = 6 mice per group for Cntl 8-week TAM-HFD, n = 7 mice per group for Trem2 ΔMФ 8-week TAM-HFD, n = 7 mice per group for Cntl 16-week TAM-HFD and n = 7 mice per group for Trem2 ΔMФ 16-week TAM-HFD). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , Confocal micrograph showing CD68 staining (green) and DAPI (blue) for macrophage area in Cntl or Trem2-deficent mice after 16-week TAM-HFD feeding. Representative image from two independent experiments. b , Quantification of CD68 + macrophage area per section in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01. c , Quantification of the percentage of plaque that is macrophages (CD68 + ) in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test. d , Confocal micrograph showing Ki67 staining (magenta) and CD68 staining (green) for proliferation in Cntl or Trem2-deficient mice after 16-week TAM-HFD feeding. Representative image from two independent experiments. e , Quantification of Ki67 + macrophages (CD68 + ) per section in 8-week or 16-week TAM-HFD samples ( n = 5 mice per group for 8-week TAM-HFD, n = 7 mice per group for Cntl 16-week TAM-HFD and n = 6 mice per group for 16-week TAM-HFD Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01. f , Confocal micrograph of TUNEL staining (magenta) and CD68 staining (green) for detection of dying cells within atherosclerotic lesions after 16-week TAM-HFD feeding. Representative image from two independent experiments. g , Quantification of TUNEL + macrophages (CD68 + ) per section in 8-week or 16-week TAM-HFD samples ( n = 6 mice per group for Cntl 8-week TAM-HFD, n = 7 mice per group for Trem2 ΔMФ 8-week TAM-HFD, n = 7 mice per group for Cntl 16-week TAM-HFD and n = 7 mice per group for Trem2 ΔMФ 16-week TAM-HFD). Data are mean ± s.e.m. Student’s t -test, * P < 0.05 and ** P < 0.01.

Techniques: Staining, TUNEL Assay

a , Schematic for intervention study where mice were fed an HFD for 8 weeks and then switched to TAM/HFD for an additional 8 weeks before being killed. b , En face aorta analysis of plaque area after 16 weeks of diet-switch intervention study. Representative image from two independent experiments. c , Quantification of plaque area in aorta and aortic sinus ( n = 8 mice per group for Cntl and n = 9 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. d , Blood immune population analysis after 16-week diet-switch intervention model ( n = 5 mice per group). Data are mean ± s.e.m. e , Total serum cholesterol levels after 16-week diet-switch model ( n = 4 mice per group for Cntl and n = 5 for Trem2 ΔMФ ). Data are mean ± s.e.m. f , Quantification of plaque macrophage proliferation analysis by Ki67 + macrophages (CD68 + ) ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. g , Quantification of TUNEL + macrophages (CD68 + ) in plaques after 16-week diet-switch model ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. ORO, Oil Red O.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , Schematic for intervention study where mice were fed an HFD for 8 weeks and then switched to TAM/HFD for an additional 8 weeks before being killed. b , En face aorta analysis of plaque area after 16 weeks of diet-switch intervention study. Representative image from two independent experiments. c , Quantification of plaque area in aorta and aortic sinus ( n = 8 mice per group for Cntl and n = 9 for Trem2 ΔMФ ). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01. d , Blood immune population analysis after 16-week diet-switch intervention model ( n = 5 mice per group). Data are mean ± s.e.m. e , Total serum cholesterol levels after 16-week diet-switch model ( n = 4 mice per group for Cntl and n = 5 for Trem2 ΔMФ ). Data are mean ± s.e.m. f , Quantification of plaque macrophage proliferation analysis by Ki67 + macrophages (CD68 + ) ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. g , Quantification of TUNEL + macrophages (CD68 + ) in plaques after 16-week diet-switch model ( n = 5 mice per group). Data are mean ± s.e.m. Student’s t -test, * P < 0.05. ORO, Oil Red O.

Techniques: TUNEL Assay

a , WT and Trem2 −/− BV2s assessed for Trem2 expression by flow cytometry. WT ( b ) or Trem2 −/− ( c ) BV2 macrophages DEGs from bulk RNA-seq determined by Wald test with DESeq2. d , GSEA plot of cholesterol biosynthesis pathways. ES, enrichment score. e , Pathway analysis of RNA-seq data comparing WT and Trem2 −/− non-foamy BV2 cells. f , Pathway analysis of RNA-seq data comparing WT and Trem2 −/− foamy BV2 cells. e , f , Significant pathways determined using weighted Kolmogorov–Smirnov test. g , WT or Trem2 −/− cell supernatant assessed for cytotoxicity by LDH assay after 16 h ( n = 6 biological replicates per group). Foamy: 20 μg ml −1 cholesterol; foamy hi : 80 μg ml −1 cholesterol. Data are mean ± s.e.m. Two-tailed ANOVA, *** P < 0.001. h , DiI-oxLDL uptake for WT or Trem2 −/− non-foamy and foamy BV2 macrophages ( n = 6 for non-foamy WT and Trem2 −/− and n = 4 foamy WT and Trem2 −/− biological replicates). Data are mean ± s.e.m. Student’s t -test, *** P < 0.001. i , WT or Trem2 −/− non-foamy and foamy BV2 macrophage efferocytosis. Efferocytotic cells were determined by the percent of BV2s that were positive for CTV-labeled splenocytes ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01 and **** P < 0.0001. j , WT or Trem2 −/− non-foamy and foamy BV2 macrophage sXBP1 expression. Tunicamycin was used as a positive control ( n = 6 biological replicates per group). FMO (fluorescence minus one) shows unstained control. Data are mean ± s.e.m. Two-tailed ANOVA, ** P < 0.01. k , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) plus 10 μM PBA. Cell supernatant was assessed for cytotoxicity by LDH assay after 16 h ( n = 6 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. l , GSEA plot of cholesterol efflux pathways from RNA-seq. m , GSEA plot of NR1H2 and NR1H3 gene target pathways from RNA-seq. n , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) ± T0901317 percent cytotoxicity ( n = 5 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. o , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) ± 10 μM T0901317, assessed for sXBP1 levels by flow cytometry. Tunicamycin was used as a positive control ( n = 5 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. KO, knockout; NS, not significant.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a , WT and Trem2 −/− BV2s assessed for Trem2 expression by flow cytometry. WT ( b ) or Trem2 −/− ( c ) BV2 macrophages DEGs from bulk RNA-seq determined by Wald test with DESeq2. d , GSEA plot of cholesterol biosynthesis pathways. ES, enrichment score. e , Pathway analysis of RNA-seq data comparing WT and Trem2 −/− non-foamy BV2 cells. f , Pathway analysis of RNA-seq data comparing WT and Trem2 −/− foamy BV2 cells. e , f , Significant pathways determined using weighted Kolmogorov–Smirnov test. g , WT or Trem2 −/− cell supernatant assessed for cytotoxicity by LDH assay after 16 h ( n = 6 biological replicates per group). Foamy: 20 μg ml −1 cholesterol; foamy hi : 80 μg ml −1 cholesterol. Data are mean ± s.e.m. Two-tailed ANOVA, *** P < 0.001. h , DiI-oxLDL uptake for WT or Trem2 −/− non-foamy and foamy BV2 macrophages ( n = 6 for non-foamy WT and Trem2 −/− and n = 4 foamy WT and Trem2 −/− biological replicates). Data are mean ± s.e.m. Student’s t -test, *** P < 0.001. i , WT or Trem2 −/− non-foamy and foamy BV2 macrophage efferocytosis. Efferocytotic cells were determined by the percent of BV2s that were positive for CTV-labeled splenocytes ( n = 5 biological replicates per group). Data are mean ± s.e.m. Student’s t -test, ** P < 0.01 and **** P < 0.0001. j , WT or Trem2 −/− non-foamy and foamy BV2 macrophage sXBP1 expression. Tunicamycin was used as a positive control ( n = 6 biological replicates per group). FMO (fluorescence minus one) shows unstained control. Data are mean ± s.e.m. Two-tailed ANOVA, ** P < 0.01. k , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) plus 10 μM PBA. Cell supernatant was assessed for cytotoxicity by LDH assay after 16 h ( n = 6 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. l , GSEA plot of cholesterol efflux pathways from RNA-seq. m , GSEA plot of NR1H2 and NR1H3 gene target pathways from RNA-seq. n , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) ± T0901317 percent cytotoxicity ( n = 5 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. o , WT or Trem2 −/− foamy BV2 macrophages (80 μg ml −1 cholesterol) ± 10 μM T0901317, assessed for sXBP1 levels by flow cytometry. Tunicamycin was used as a positive control ( n = 5 biological replicates per group). Data are mean ± s.e.m. Two-tailed ANOVA, **** P < 0.0001. KO, knockout; NS, not significant.

Techniques: Expressing, Flow Cytometry, RNA Sequencing, Lactate Dehydrogenase Assay, Two Tailed Test, Labeling, Positive Control, Fluorescence, Control, Knock-Out

a ) Heat map of nonfoamy macrophages comparing top enriched WT and Trem2−/− genes. b ) Heat map of foamy macrophages comparing top enriched WT and Trem2−/− genes. c ) Normalized enrichment scores for top pathways associated with WT BV2 compared to Trem2−/− BV2 in media alone (nonfoamy) or following foamy differentiation. Significant pathways were determined using Weighted-Kolmogorov-Smirnov (WKS) test. d ) Heat map of nonfoamy and foamy macrophages comparing matrix metalloprotease genes from WT and Trem2−/− BV2. e ) Cytokine supernatant analysis from cultured WT or Trem2−/− cells cultured with either media alone or media with 20 mg/ml soluble cholesterol overnight (n = 3 biological replicates/group). Data are mean ±S.E.M.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) Heat map of nonfoamy macrophages comparing top enriched WT and Trem2−/− genes. b ) Heat map of foamy macrophages comparing top enriched WT and Trem2−/− genes. c ) Normalized enrichment scores for top pathways associated with WT BV2 compared to Trem2−/− BV2 in media alone (nonfoamy) or following foamy differentiation. Significant pathways were determined using Weighted-Kolmogorov-Smirnov (WKS) test. d ) Heat map of nonfoamy and foamy macrophages comparing matrix metalloprotease genes from WT and Trem2−/− BV2. e ) Cytokine supernatant analysis from cultured WT or Trem2−/− cells cultured with either media alone or media with 20 mg/ml soluble cholesterol overnight (n = 3 biological replicates/group). Data are mean ±S.E.M.

Techniques: Cell Culture

a ) WT or Trem2−/− peritoneal macrophages were differentiated in media control, media with 20 μg/mL or 80 μg/mL soluble cholesterol to induce foamy macrophage formation. Cell supernatant was assessed for cytotoxicity by LDH assay after 16 hours (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = * < 0.05. b ) WT or Trem2−/− peritoneal macrophages were differentiated in media control or media with 20 μg/mL soluble cholesterol, then cultured with irradiated, cell trace violet (CTV) labeled splenocytes for 2 hours. Percentage of efferocytotic cells were determined by the % of peritoneal macrophages that were positive for CTV labeled splenocytes (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = *** < 0.001, ****<0.0001. c ) WT or Trem2−/− peritoneal macrophages were differentiated in media control or media with 20 μg/mL soluble cholesterol, then assessed for activation of ER stress response by sXBP1 levels by flow cytometry. Tunicamycin was used as a positive control (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = *** < 0.001.

Journal: Nature cardiovascular research

Article Title: Trem2 promotes foamy macrophage lipid uptake and survival in atherosclerosis

doi: 10.1038/s44161-023-00354-3

Figure Lengend Snippet: a ) WT or Trem2−/− peritoneal macrophages were differentiated in media control, media with 20 μg/mL or 80 μg/mL soluble cholesterol to induce foamy macrophage formation. Cell supernatant was assessed for cytotoxicity by LDH assay after 16 hours (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = * < 0.05. b ) WT or Trem2−/− peritoneal macrophages were differentiated in media control or media with 20 μg/mL soluble cholesterol, then cultured with irradiated, cell trace violet (CTV) labeled splenocytes for 2 hours. Percentage of efferocytotic cells were determined by the % of peritoneal macrophages that were positive for CTV labeled splenocytes (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = *** < 0.001, ****<0.0001. c ) WT or Trem2−/− peritoneal macrophages were differentiated in media control or media with 20 μg/mL soluble cholesterol, then assessed for activation of ER stress response by sXBP1 levels by flow cytometry. Tunicamycin was used as a positive control (n = 5 biological replicates/group). Data are mean ± S.E.M. Two-tailed ANOVA, P = *** < 0.001.

Techniques: Control, Lactate Dehydrogenase Assay, Two Tailed Test, Cell Culture, Irradiation, Labeling, Activation Assay, Flow Cytometry, Positive Control

Figure 2. Gating strategy for FACS-sorting of wound macrophages (A and B) Representative ﬂow cytometry plots at (A) 4 and (B) 14 days post injury. DAPI- single cells were gated (Gates G1-G4) and expression of CD45 was analyzed. CD45+ cells were gated (G5) and analyzed for expression of CD11b and F4/80. Single CD45+CD11b+F4/80+ wound macrophages (G6) were sorted in 384-well plates for RNA sequencing. CD45+CD11b+F4/80+ wound macrophages can further be analyzed for expression of Trem2, CD301b, and MHC II. Index sorting allows merging FACS data with gene expression data at single cell level.

Journal: STAR protocols

Article Title: Isolation of macrophages from mouse skin wounds for single-cell RNA sequencing.

doi: 10.1016/j.xpro.2022.101337

Figure Lengend Snippet: Figure 2. Gating strategy for FACS-sorting of wound macrophages (A and B) Representative ﬂow cytometry plots at (A) 4 and (B) 14 days post injury. DAPI- single cells were gated (Gates G1-G4) and expression of CD45 was analyzed. CD45+ cells were gated (G5) and analyzed for expression of CD11b and F4/80. Single CD45+CD11b+F4/80+ wound macrophages (G6) were sorted in 384-well plates for RNA sequencing. CD45+CD11b+F4/80+ wound macrophages can further be analyzed for expression of Trem2, CD301b, and MHC II. Index sorting allows merging FACS data with gene expression data at single cell level.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies APC Rat anti-mouse Trem2 (237920), 100 tests, dilution 1:10 R&D Systems Cat# FAB17291A; RRID: AB_884527 APC-eFluor780 Rat anti-mouse CD11b (M1/70), 0.2 mg/mL, dilution 1:400 Thermo Fisher Scientific Cat# 47-0112-80; RRID: AB_1603195 eFluor450 Rat anti-mouse MHC II (M5/ 114.15.2), 0.2 mg/mL, dilution 1:200 Thermo Fisher Scientific Cat# 48-5321-82; RRID: AB_1272204 FITC Rat anti-mouse CD45 (30-F11), 0.2 mg/mL, dilution 1:200 Thermo Fisher Scientific Cat# 11-0451-82; RRID: AB_465050 PE Rat anti-mouse F4/80 (CI:A3-1), 100 tests, dilution 1:10 Bio-Rad Laboratories Cat# MCA497PE; RRID: 2098196 PerCP/Cy5.5 Rat anti mouse CD301b (URA-1), 0.2 mg/mL, dilution 1:50 BioLegend Cat# 146809; RRID: AB_2563391 Rat anti-mouse CD16/CD32, 0.5 mg/mL, dilution 1:50 Thermo Fisher Scientific Cat# 14-0161-82; RRID: AB_467133 Chemicals, peptides, and recombinant proteins Betaine (5 M) Thermo Fisher Scientific Cat# J77507.AB (Continued on next page) 2 STAR Protocols 3, 101337, June 17, 2022 Continued REAGENT or RESOURCE SOURCE IDENTIFIER dNTP Thermo Fisher Scientific (Life Technologies) Cat# R0192 EDTA (0.5 M, pH 8.0) solution Thermo Fisher Scientific (Ambion) Cat# AM9261 Ethanol Merck Cat# 1009835000 H2O, sterile Fresenius Kabi 40676.00.00 Liberase TM Research Grade Roche Cat# 05401127001 MgCl2 (1 M) Thermo Fisher Scientific (Invitrogen) Cat# AM9530G Murine RNase Inhibitor New England Biolabs Cat# M0314 S NaCl (5 M) Thermo Fisher Scientific (Invitrogen) Cat# AM9759 Poly(ethylene glycol) avg mol wt 8,000 Sigma-Aldrich Cat# P5413-500G Red Blood Cell Lysis Solution Miltenyi Biotec Cat# 130-094-183 TE buffer (13) Thermo Fisher Scientific (Invitrogen) Cat# AM9858 Tris-Buffer pH 8.0 (1 M) VWR (AppliChem) Cat# A4577.1000 Triton-X 100 Merck Cat# T8787-50ML Critical commercial assays AccuBlue Broad Range ds DNA Kit, incl 9 standards VWR/Biotium Cat# 31007 BD CaliBRITE beads BD Biosciences Cat# 340486 ERCC spike-in Thermo Fisher Scientific (Invitrogen/Ambion) Cat# 4456740 HS NGS Fragment Kit (1–6,000 bp) Agilent Cat# NF-474-0500 Illumina DNA Prep, (M) Tagmentation (96 Samples) Illumina Cat# 20018705 Kapa HiFi HotStart Readymix (23) Roche Cat# 7958935001 Magnetic beads (GE; Sera-Mag SpeedBead Carboxylate-Modified Magnetic Particles (Hydrophobic)) GE Healthcare Cat# 65152105050250 SuperScript II Reverse Transcriptase, incl.

Techniques: Cytometry, Expressing, RNA Sequencing, Gene Expression

Review

flow n a trem2 apc 237920 rat igg2b r d systems fab17291a (R&D Systems)

Structured Review

Images

Related Articles

Expressing:

CRISPR:

Knock-Out:

Incubation:

Isolation:

Flow Cytometry:

Staining:

Cell Culture:

Control:

Immunohistochemical staining:

Irradiation:

Derivative Assay:

Labeling:

Injection:

Multiplex Assay:

Immunofluorescence:

TUNEL Assay:

RNA Sequencing:

Lactate Dehydrogenase Assay:

Two Tailed Test:

Positive Control:

Fluorescence:

Activation Assay:

Similar Products

Image Search Results