Journal: Frontiers in Cellular Neuroscience

Article Title: TREM2 Regulates Heat Acclimation-Induced Microglial M2 Polarization Involving the PI3K-Akt Pathway Following EMF Exposure

doi: 10.3389/fncel.2019.00591

Figure Lengend Snippet: Effects of HA on the expression of triggering receptor expressed on myeloid cells-2 (TREM2) and the phosphorylation of PI3K and Akt in EMF-stimulated N9 cells. N9 cells were pretreated with or without a 72-h HA process (20/4-h cycle of 37°C/39.5°C) and then exposed to 2.45-GHz EMF or sham-exposed for 20 min. Levels of TREM2 (A) , and phosphorylation of PI3K and Akt (B) in total cell lysates were analyzed using Western blotting, and the corresponding densitometric analyses were represented. Data are presented as means ± SEM of three independent experiments. * P < 0.05 vs. the sham-exposed control group; # P < 0.05 vs. the EMF-exposed group.

Article Snippet: The membranes were blocked in PBS with 5% non-fat milk for 1 h and then incubated with their respective primary antibodies against rat anti-mouse CD11b (1:800, AbD Serotec, Oxford, UK), mouse anti-mouse CD86 (1:800, Abcam, Cambridge, MA, USA), and rat anti-mouse TREM2 (1:500, Merck Millipore, Temecula, CA, USA), and with antibodies purchased from Santa-Cruz Biotechnology (Santa Cruz, USA) that recognize mouse anti-mouse CD206 (1:100) and mouse anti-mouse Arg1 (1:100), and from Cell Signaling Technology (Danvers, MA, USA) that recognize rabbit anti-mouse phosphor-PI3K p85 Tyr458 (p-PI3K, 1:800), rabbit anti-mouse phospho-Akt Ser473 (p-Akt, 1:1,000).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control

Journal: Frontiers in Cellular Neuroscience

Article Title: TREM2 Regulates Heat Acclimation-Induced Microglial M2 Polarization Involving the PI3K-Akt Pathway Following EMF Exposure

doi: 10.3389/fncel.2019.00591

Figure Lengend Snippet: Effects of TREM2 esiRNA on M2 microglial phenotype regulation in EMF-stimulated N9 cells with HA preconditioning. N9 cells were transfected with or without TREM2 esiRNA (20 nM) at 60 h during the uncompleted 72-h HA and then continuously cultured 12 h to complete the HA process and the 24 h transfection of TREM2 esiRNA. Western blotting quantification of TREM2 (A) , and M2 markers CD206 and Arg1 (C) , and ELISA of anti-inflammatory cytokines IL-4 and IL-10 (B) production of in either control or HA-plus-EMF-treated N9 cells with or without TREM2 esiRNA. Data are presented as means ± SEM of three independent experiments. * P < 0.05 vs. the non-HA and sham-exposed control group; # P < 0.05 vs. the HA-plus-EMF-exposed group. (D) Confocal immunofluorescence microscopy was performed on cultures that were immunoreacted with antibodies against TREM2 and CD206 with esi-TREM2 treatment in HA-plus-EMF-treated N9 cells. Scale bar: 20 μm.

Article Snippet: The membranes were blocked in PBS with 5% non-fat milk for 1 h and then incubated with their respective primary antibodies against rat anti-mouse CD11b (1:800, AbD Serotec, Oxford, UK), mouse anti-mouse CD86 (1:800, Abcam, Cambridge, MA, USA), and rat anti-mouse TREM2 (1:500, Merck Millipore, Temecula, CA, USA), and with antibodies purchased from Santa-Cruz Biotechnology (Santa Cruz, USA) that recognize mouse anti-mouse CD206 (1:100) and mouse anti-mouse Arg1 (1:100), and from Cell Signaling Technology (Danvers, MA, USA) that recognize rabbit anti-mouse phosphor-PI3K p85 Tyr458 (p-PI3K, 1:800), rabbit anti-mouse phospho-Akt Ser473 (p-Akt, 1:1,000).

Techniques: esiRNA, Transfection, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Control, Immunofluorescence, Microscopy

Journal: Frontiers in Cellular Neuroscience

Article Title: TREM2 Regulates Heat Acclimation-Induced Microglial M2 Polarization Involving the PI3K-Akt Pathway Following EMF Exposure

doi: 10.3389/fncel.2019.00591

Figure Lengend Snippet: Effects of TREM2 esiRNA and the PI3K inhibitor LY294002 on the phosphorylation of PI3K and Akt in EMF-stimulated N9 cells with HA preconditioning. N9 cells were transfected with or without TREM2 esiRNA (20 nM) at 60 h during the uncompleted 72-h HA and then continuously cultured 12 h to complete the HA process and the 24 h transfection of TREM2 esiRNA. Alternatively, N9 cells were treated with or without LY294002 (10 μM) for 30 min prior to the end of the 72-h HA process. Levels of PI3K and Akt phosphorylation in total cell lysates were analyzed using Western blotting, and the corresponding densitometric analyses were represented. Data are presented as means ± SEM of three independent experiments. * P < 0.05 vs. the non-HA and sham-exposed control group; # P < 0.05 vs. the HA-plus-EMF-exposed group.

Article Snippet: The membranes were blocked in PBS with 5% non-fat milk for 1 h and then incubated with their respective primary antibodies against rat anti-mouse CD11b (1:800, AbD Serotec, Oxford, UK), mouse anti-mouse CD86 (1:800, Abcam, Cambridge, MA, USA), and rat anti-mouse TREM2 (1:500, Merck Millipore, Temecula, CA, USA), and with antibodies purchased from Santa-Cruz Biotechnology (Santa Cruz, USA) that recognize mouse anti-mouse CD206 (1:100) and mouse anti-mouse Arg1 (1:100), and from Cell Signaling Technology (Danvers, MA, USA) that recognize rabbit anti-mouse phosphor-PI3K p85 Tyr458 (p-PI3K, 1:800), rabbit anti-mouse phospho-Akt Ser473 (p-Akt, 1:1,000).

Techniques: esiRNA, Phospho-proteomics, Transfection, Cell Culture, Western Blot, Control

Journal: Alzheimer's & Dementia : Translational Research & Clinical Interventions

Article Title: Alzheimer's disease drug development pipeline: 2021

doi: 10.1002/trc2.12179

Figure Lengend Snippet: Agents in Phase 2 of Alzheimer's disease drug development (ClinicalTrials.gov accessed January 5, 2021)

Article Snippet: AL002 , Inflammation , Monoclonal antibody targeting TREM2 receptors to promote microglial clearance of Aβ , DMT , Recruiting (NCT04592874) , Alector, AbbVie , Nov 2020 , Aug 2023.

Techniques: Infection, Activity Assay, Binding Assay, Expressing, Reverse Transcription, Phospho-proteomics

Journal: Cell reports

Article Title: Comparing neoantigen cancer vaccines and immune checkpoint therapy unveils an effective vaccine and anti-TREM2 macrophage-targeting dual therapy

doi: 10.1016/j.celrep.2024.114875

Figure Lengend Snippet: (A) UMAP displaying subclustering of select myeloid clusters from CD45 + scRNA-seq and heatmap displaying normalized expression of select genes by monocyte/macrophage cluster. (B) Bar graphs depicting frequency of monocytes/macrophages in each cluster by treatment. (C) Heatmap displaying normalized expression of Mrc1 (CD206), Cx3cr1 , and Nos2 (iNOS) in each monocyte/macrophage cluster by treatment. (D) scRNA-seq dot plot depicting Trem2 and Cx3cr1 expression in combined monocyte/macrophage clusters. (E and F) Representative flow cytometry plots and graphs displaying intratumoral (E) CX3CR1 + CD206 + macrophages or (F) iNOS + macrophages from Y1.7 melanoma-bearing mice treated beginning on day 7 post-tumor transplant and harvested on day 15. For (E) and (F), dot plots displaying CX3CR1 + CD206 + and iNOS + macrophages are gated on macrophages, and bar graphs display the mean ± SEM and are representative of at least three independent experiments (* p < 0.05, ** p < 0.01, **** p < 0.0001; NS, not significant, unpaired t test). See also and .

Article Snippet: Anti-Mouse TREM2 - Fc Muted [clone 178 (LALAPG)] , Leinco Technologies , Cat# T721.

Techniques: Expressing, Flow Cytometry

Journal: Cell reports

Article Title: Comparing neoantigen cancer vaccines and immune checkpoint therapy unveils an effective vaccine and anti-TREM2 macrophage-targeting dual therapy

doi: 10.1016/j.celrep.2024.114875

Figure Lengend Snippet: (A) Trem2 mRNA detection by quantitative reverse-transcriptase PCR (qRT-PCR) on sorted intratumoral CX3CR1 + CD206 + macrophages and non-CX3CR1 + CD206 + macrophages isolated on day 19 post-tumor transplant from Y1.7LI tumor-bearing mice treated with neoAg SLP vax on days 12 and 18. (B) Schematic depicting the experiment in (C). (C) Tumor growth in mice transplanted with Y1.7LI melanoma cells and receiving intratumoral injections of CX3CR1 + CD206 + macrophages or PBS and treated with control vax, neoAg SLP vax, anti-TREM2, neoAg SLP vax + isotype control mAb (Iso), or neoAg SLP vax + anti-TREM2 as indicated in (B). (D) Tumor growth in Y1.7 melanoma-bearing mice treated with Iso, anti-TREM2, Iso + control vax, Iso + neoAg SLP vax, anti-TREM2 + control vax, or anti-TREM2 + neoAg SLP vax. (E) Schematic depicting experiments in (F)–(H). (F) Graphs displaying frequency of intratumoral CX3CR1 + CD206 + macrophages and iNOS + macrophages. (G) Graph displaying frequency of mLama4 tetramer-positive CD8 T cells. (H) Graph displaying IFN-g + CD8 T cells as assessed by ICS of mLama4 peptide-restimulated CD8 T cells isolated from Y1.7LI tumors. For (A), RNA was isolated from macrophages from two individual mice (two independent experiments). For (C) and (D), fractions indicate number of mice rejecting tumors/number of mice used in the experiment. Scatterplots in (F)–(H) display data for individual mice and are representative of two independent experiments (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; NS, not significant, unpaired t test).

Article Snippet: Anti-Mouse TREM2 - Fc Muted [clone 178 (LALAPG)] , Leinco Technologies , Cat# T721.

Techniques: Reverse Transcription, Quantitative RT-PCR, Isolation, Control

Journal: Cell reports

Article Title: Comparing neoantigen cancer vaccines and immune checkpoint therapy unveils an effective vaccine and anti-TREM2 macrophage-targeting dual therapy

doi: 10.1016/j.celrep.2024.114875

Figure Lengend Snippet:

Article Snippet: Anti-Mouse TREM2 - Fc Muted [clone 178 (LALAPG)] , Leinco Technologies , Cat# T721.

Techniques: Control, Virus, Recombinant, Lysis, Cloning, Mutagenesis, Sequencing, SYBR Green Assay, Staining, dsDNA Assay, Library Quantification, Software, Real-time Polymerase Chain Reaction