Journal: bioRxiv

Article Title: YAP/TEAD drives treatment-induced adaptive immunosuppression in EGFR-mutant lung cancer

doi: 10.64898/2026.01.22.701073

Figure Lengend Snippet: A) GSEA analysis of MoMacVERSE monocyte/macrophage signatures using RNA-sequencing data from . B) GSEA enrichment plots of two independent TREM2 + macrophage signatures , . C) GSEA of Hallmark genes. D) Flow cytometry analysis of TREM2 surface expression in monocyte-derived macrophages exposed to HCC827 CM. E) Immunohistochemical staining of TREM2 from syngeneic Del19.2 model in vivo experiment depicted in . F) Quantification of TREM2 + area from samples in (E) using QuPath. G) TREM2 gene expression in EGFR TKI-treated patients. RNA-sequencing data from Gurule et al. . H) Correlation of macrophage and T cell abundance from CIBERSORTx analysis of RNA sequencing data in (G). CM = conditioned medium. Paired (D) or unpaired (F) t-tests were used to assess statistical significance. **, P-value <0.01; *, P-value < 0.05.

Article Snippet: Surface staining for markers was carried out in PBS + 2 % FBS with the following antibodies (1:100 dilution): CD64-PE (clone 10.2, BD PharmingenTM), CD206-AF488 (clone 15-4, BioLegend), CD163-BV711 (clone GHI_61, BD HorizonTM), and TREM2-APC (clone 237920, R&D Systems).

Techniques: RNA Sequencing, Flow Cytometry, Expressing, Derivative Assay, Immunohistochemical staining, Staining, In Vivo, Gene Expression

Journal: bioRxiv

Article Title: YAP/TEAD drives treatment-induced adaptive immunosuppression in EGFR-mutant lung cancer

doi: 10.64898/2026.01.22.701073

Figure Lengend Snippet: A) Experimental set-up for conditioned media collection and YAP/TEAD inhibition in DTP cells. B) RNA-sequencing analysis of the expression of genes encoding for secreted proteins in HCC827 and HCC4006 cell lines treated as in (A). C) YAP-driven secretome in HCC827 and HCC4006 cells, determined by genes upregulated in DTPs, and downregulated upon TEAD inhibition. D) Shared YAP-driven secretome in HCC827 and HCC4006. E) ChIP-seq analysis of YAP binding of shared secretome genes. F) List of shared YAP-driven secretome from (D), with genes previously associated with macrophage activation, migration, or polarization highlighted in bold. G) CD163 expression of THP-1 cells treated with HCC827 conditioned media collected as in (A) using 500 nM ORM-47286 or 10 mM MYF-01-37 for TEAD inhibition. H) Flow cytometry analysis of CD163 and TREM2 cell surface expression in human peripheral blood monocytes exposed to HCC827 conditioned media. Data in (G) presented as mean ± SD. Unpaired (G) and paired (H) one-way ANOVA was used to assess statistical significance. **, P-value <0.01; *, P-value < 0.05.

Article Snippet: Surface staining for markers was carried out in PBS + 2 % FBS with the following antibodies (1:100 dilution): CD64-PE (clone 10.2, BD PharmingenTM), CD206-AF488 (clone 15-4, BioLegend), CD163-BV711 (clone GHI_61, BD HorizonTM), and TREM2-APC (clone 237920, R&D Systems).

Techniques: Inhibition, RNA Sequencing, Expressing, ChIP-sequencing, Binding Assay, Activation Assay, Migration, Flow Cytometry

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: High-fat diet feeding increases cleaved TREM2 and TREM2 levels in gonadal white adipose tissue (A) Immunoblot analysis of TREM2 and cleaved TREM2 levels in gonadal white adipose tissue (GWAT), inguinal white adipose tissue (IWAT), and brown adipose tissue (BAT) of mice fed a high-fat diet (HFD) and normal chow diet (NCD) for 4 and 8 weeks ( n = 6 mice per group). (B) ELISA-based quantification of soluble TREM2 (sTREM2) levels in mouse serum ( n = 3 mice per group). (C) Schematic diagram illustrating cleaved membrane-bound TREM2 and soluble TREM2 forms. Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Membrane

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: HFD feeding upregulates Adam10 and Adam17 expression in monocyte/macrophage populations of GWAT (A) Immunoblot analysis of ADAM10 and ADAM17 protein expression in gonadal white adipose tissue (GWAT) of mice after 8 weeks of NCD or HFD feeding ( n = 6 mice per group). (B) Umap plot of Adam10 , Adam17 , and total nuclei isolated from GWAT of NCD and HFD-fed mice (PRJNA942977). Clusters are colored by cell types: adipocyte, mesothelial cell, lymphatic endothelial cell, vascular endothelial cell, adipocyte progenitor cell, smooth muscle cell (SMC), monocyte/macrophage (mono/mac), dendritic cell (DC), B cell, and T cell. (C) Violin plot of Adam10 and Adam17 gene expression levels in total cell types from GWAT of NCD- and HFD-fed mice. (D) Umap plot of Adam10 , Adam17 from monocyte/macrophage population in GWAT of NCD- and HFD-fed mice (PRJNA942977). Clusters are colored by cell types: Lyve1 +macrophage (Mac. Lyve1 ), Trem2 +macrophage (Mac. Trem2 ), Prg4 +macrophage (Mac. Prg4 ), and monocyte. (E) Bubble plot and violin plot of Adam10 and Adam17 expression levels in distinct cell populations from mouse GWAT, determined by single-nucleus RNA sequencing (PRJNA942977). (F) qPCR analysis of Adam10 and Adam17 mRNA expression in isolated F4/80+ macrophages and adipocytes from GWAT after 8 weeks of NCD or HFD feeding ( n = 3 mice per group). Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Expressing, Western Blot, Isolation, Gene Expression, RNA Sequencing

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: Obesity increases ADAM10 and ADAM17 expression in macrophage populations of WAT (A) ADAM10 and ADAM17 expression levels in total cell types of human visceral adipose tissues ( GSE176171 ). Clusters are colored by cell types: Adipocyte, adipose stem and progenitor cells (ASPC), mesothelium, endothelial cell, lymphatic endothelial cell (LEC), pericyte, smooth muscle cell (SMC), macrophage, monocyte, dendritic cell (DC), mast cell, neutrophil, B cell, natural killer cell (NK cell), T cell, and endometrium cell. (B) Bubble plot and violin plot of ADAM10 and ADAM17 gene expressions and representative markers in adipocyte and macrophage populations with body mass index (BMI) in human subcutaneous adipose tissue ( GSE176171 ). (C) Violin plot of TREM2 , ADAM10 , and ADAM17 gene expression levels in human macrophage sub-populations (hMac1, hMac2, hMac3). (D) Correlation analysis of ADAM10 and ADAM17 gene expression levels with BMI in human subcutaneous adipose tissue ( n = 12 patients per group). Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Expressing, Gene Expression

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: Apoptotic adipocytes fail to induce TREM2 shedding in RAW264.7 cells and bone marrow-derived macrophages (BMDMs) (A) Schematic diagram illustrating the experimental method used for co-culturing dying/dead adipocytes with RAW264.7 cells or BMDMs. Apoptotic adipocytes (aAC) were generated by treating differentiated 3T3-L1 adipocytes with brefeldin A (BFA; 5 μg/mL for 24 h). Apoptotic adipocytes (1 × 10 5 cells/well) were directly co-cultured with RAW264.7 cells or BMDMs (5 × 10 5 cells/well) in growth medium. After 24 h of co-culture, non-engulfed or floating apoptotic adipocytes were removed by PBS washing, and RAW264.7 cells or BMDMs were subsequently harvested for immunoblot analysis. (B) Immunoblot analysis of caspase-3 expression levels in 3T3-L1 adipocytes after BFA for 24 h ( n = 3 cells per group). (C) and (D) Immunoblot analysis of TREM2, ADAM10, and ADAM17 protein expression levels in RAW264.7 cells (C) and BMDMs (D) co-cultured with apoptotic adipocytes ( n = 3 cells per group). Black arrow and red arrow indicate each precursor form of ADAM10/17 and active form of ADAM10/17. Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Derivative Assay, Generated, Cell Culture, Co-Culture Assay, Western Blot, Expressing

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: GM6001 blocks TREM2 shedding in RAW264.7 cells induced by pyroptotic adipocytes (A) A schematic diagram illustrating the experimental method used for co-culturing pyroptotic adipocytes with macrophages. Pyroptotic adipocytes were generated by treating differentiated 3T3L1 adipocytes with lipopolysaccharide (LPS; 100 μg/μL, 48 h) followed by ATP (2 mM, 24 h). Pyroptotic adipocytes (1 × 10 5 cells/well) were directly co-cultured with RAW264.7 cells (5 × 10 5 cells/well) or BMDMs for 24 h in growth medium. After co-culture, non-engulfed or floating pyroptotic adipocytes were removed by PBS washing, and RAW264.7 cells and BMDMs were subsequently harvested for immunoblot analysis. (B) Immunoblot analysis of NLRP3, caspase-1, and GSDMD (gasdermin D) expression levels in 3T3-L1 adipocytes after LPS priming for 48 h and ATP treatment for 24 h ( n = 3 cells per group). Black arrow and red arrow indicate each total form of GSDMD and cleaved GSDMD. (C) Immunoblot analysis of TREM2, ADAM10, and ADAM17 expression levels in RAW264.7 cells co-cultured with pyroptotic adipocytes for 24 h in the presence or absence of GM6001 ( n = 4 cells per group). Black arrow and red arrow indicate each precursor form of ADAM10/17 and active form of ADAM10/17. (D) Immunoblot analysis of P-STING and STING expression levels in RAW264.7 cells co-cultured with pyroptotic adipocytes ( n = 3 cells per group). (E) Immunoblot analysis of P-syk, syk, P-PI3K, PI3K, P-PLCγ1, PLCγ1, P-AKT, and AKT protein levels in RAW264.7 cells co-cultured with pyroptotic adipocytes ( n = 3 cells per group). (F) Phagocytosis analysis of RAW264.7 cells co-cultured with pyroptotic adipocytes for 18 h in the presence of GM6001. Adipocytes were tagged with C12-BODIPY (red), and macrophages were stained with DiO (green). Representative images from three independent experiments are shown, with quantification provided in the right panel. Scale bars, 100 μm. Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Generated, Cell Culture, Co-Culture Assay, Western Blot, Expressing, Staining

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: GM6001 prevents the cleavage of TREM2 on BMDMs induced by pyroptotic adipocytes (A) Immunoblot analysis of TREM2, ADAM10, and ADAM17 expression levels in BMDMs co-cultured with pyroptotic adipocytes for 24 h in the presence or absence of GM6001 ( n = 3 cells per group). Black arrow and red arrow indicate each precursor form of ADAM10/17 and active form of ADAM10/17. (B) and (C) Phagocytosis analysis of BMDMs, (B) TREM2 knockdown BMDMs, and negative control (NC). (C) co-cultured with pyroptotic adipocytes for 18 h in the presence of GM6001. Adipocytes were tagged with C12-BODIPY (red), and macrophages were stained with DiO (green). Representative images from three independent experiments are shown, with quantification provided in the right panel. The yellow boxes indicate the regions shown in the magnified images. Scale bars, 200 μm. Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Western Blot, Expressing, Cell Culture, Knockdown, Negative Control, Staining

Journal: iScience

Article Title: Obesity-induced pyroptotic adipocyte death leads to TREM2-dependent macrophage dysfunction and adipose tissue inflammation

doi: 10.1016/j.isci.2025.114358

Figure Lengend Snippet: GM6001 treatment reduces TREM2 shedding and HFD-induced inflammation in GWAT (A) Schematic illustration of the experimental strategy for GM6001 administration (7.5 mg/kg/2 days) in mice fed a high-fat diet (HFD) for 10 weeks. (B) Immunoblot analysis of TREM2, ADAM10, and ADAM17 protein levels in gonadal white adipose tissue (GWAT) of HFD-fed mice treated with or without GM6001 ( n = 4 mice per group). (C) Measurement of soluble TREM2 (sTREM2) levels in the serum of NCD- or HFD-fed mice treated with GM6001 for 10 weeks ( n = 3 mice per group). (D–F) Representative flow profile and quantification of (D) CD11B + CD45 + cells (E) M2/M1 macrophage ratio (CD206 + CD11C − CD45 + CD11B + /CD11C + CD206 − CD45 + CD11B + ), and (F) TREM2 + CD11C + and TREM2 + CD206 + macrophage populations in GWAT of GM6001-treated mice after feeding HFD for 10 weeks ( n = 4 mice per group). (G) Immunofluorescence staining of F4/80 (green) with DAPI (blue) counterstaining in paraffin-embedded GWAT sections from GM6001-treated and control mice ( n = 4 mice per group, scale bars, 100 μm). (H) Immunoblot analysis of P-STING, STING, NLRP3, F4/80, and caspase-1 expression levels in GWAT of HFD-fed mice treated with or without GM6001 ( n = 4 mice per group). Data are presented as mean values ± SEM. p values were determined by the unpaired two-sided Student’s t test and were annotated directly in the figure at the corresponding comparisons.

Article Snippet: Human/Mouse TREM2 Allophycocyanin Mab (FAB17291A, 1:50) were purchased from R&D systems.

Techniques: Western Blot, Immunofluorescence, Staining, Control, Expressing

Journal: Bio-protocol

Article Title: Identification and Sorting of Adipose Inflammatory and Metabolically Activated Macrophages in Diet-Induced Obesity

doi: 10.21769/BioProtoc.5479

Figure Lengend Snippet: Representative plots illustrate the sequential gating of SVF cells, including singlet and CD45 populations, macrophages (CD11bF4/80), and downstream subsets (CD11c, CD206, CD9, TREM2). Gates are indicated to show the strategy used for analysis and sorting. This figure is intended as a schematic overview of the gating hierarchy; detailed quantitative data and subset percentages are reported in Wu et al. [16].

Article Snippet: Anti-mouse Trem2 APC (R&D Systems, catalog number: FAB17291N) 25.

Techniques: