Journal: Alzheimer's Research & Therapy

Article Title: Selective reduction of ADAM10 in brain and cerebrospinal fluid of Alzheimer’s disease patients

doi: 10.1186/s13195-026-02007-6

Figure Lengend Snippet: Protein levels of ADAM10 species and mRNA in AD brain samples. Non-dementia controls (NDC, n = 13) and AD extracts ( n = 16) from prefrontal cortex were resolved by SDS-PAGE/electrophoresis prior to western blot assay. Each individual ADAM10 immunoreactive band was quantified, and levels normalized using GAPDH. A Representative western blot of ADAM10 using the rabbit anti- C-terminal region monoclonal antibody (ab124695, Abcam) and GAPDH with monoclonal antibody (60004-1-Ig, Proteintech) ( B ) Densitometric quantification of ADAM10 immunoreactive bands assigned to immature (iADAM10) and C mature (mADAM10) species normalized with respect to GAPDH. D Ratio mADAM10/iADAM10 that represents the amount of mature species vs. immature. E Relative mRNA levels of ADAM10 in NDC vs. AD patients’ samples were analyzed by qRT-PCR. Transcript levels were calculated by the comparative 2 − ΔCt method with respect to GAPDH. The graphs represent mean ± SEM. Significant P < 0.05 values assayed by t-test are indicated

Article Snippet: Representative blots of CSF samples of non-AD control individuals with large age amplitude (the age of the subjects are shown on top) probed with ( A ) anti-ADAM10 polyclonal antibody to ectodomain region (OAGA02442, Aviva) and ( B ) ADAM17 antibody towards the ectodomain region (AF9301, R&D Systems).

Techniques: SDS Page, Electrophoresis, Western Blot, Quantitative RT-PCR

Journal: Alzheimer's Research & Therapy

Article Title: Selective reduction of ADAM10 in brain and cerebrospinal fluid of Alzheimer’s disease patients

doi: 10.1186/s13195-026-02007-6

Figure Lengend Snippet: Aβ affects ADAM10 levels, but not ADAM17, in SH-SY5Y-differentiated neurons. SH-SY5Y cell cultures were differentiated to neurons with 10 µM retinoic acid treatment and then treated with 3 µM of Aβ42 for 48 h. A Representative western blot of control ( C ) and Aβ-treated (Aβ42) cell samples that was resolved with the anti C-terminal ADAM10 antibody (ab124695, Abcam). B Densitometric quantification of immunoreactive bands of iADAM10 and C mADAM10 with respect to GAPDH. Values represent the percentage with respect to control. D Values of the ratio between mature vs. immature species of ADAM10 (mADAM10/iADAM10). E Representative western blot of ADAM17 species resolved with anti-ectodomain region ADAM17 antibody (AF9301, R&D Systems) and the respective quantifications of F iADAM17 and G mADAM17 immunoreactive bands normalized to the ubiquitous protein GAPDH. H Result of the ratio of mADAM17/iADAM17. The graphs represent mean ± SEM of n = 12 samples of 3 independent experiments. Significant P < 0.05 values assayed by t-test are indicated

Article Snippet: Representative blots of CSF samples of non-AD control individuals with large age amplitude (the age of the subjects are shown on top) probed with ( A ) anti-ADAM10 polyclonal antibody to ectodomain region (OAGA02442, Aviva) and ( B ) ADAM17 antibody towards the ectodomain region (AF9301, R&D Systems).

Techniques: Western Blot, Control

Journal: Alzheimer's Research & Therapy

Article Title: Selective reduction of ADAM10 in brain and cerebrospinal fluid of Alzheimer’s disease patients

doi: 10.1186/s13195-026-02007-6

Figure Lengend Snippet: ADAM10 and ADAM17 CSF levels are not altered with aging. Representative blots of CSF samples of non-AD control individuals with large age amplitude (the age of the subjects are shown on top) probed with ( A ) anti-ADAM10 polyclonal antibody to ectodomain region (OAGA02442, Aviva) and ( B ) ADAM17 antibody towards the ectodomain region (AF9301, R&D Systems). Correlations between the age and the levels of ADAM10 and ADAM17 species ( C ) iADAM10, ( D ) mADAM10, ( E ) sADAM10, ( F ) iADAM17, ( G ) mADAM17 and ( H ) sADAM17 were assayed. All graphs include their Spearman coefficient (r) and the P value. None of the ADAM10 species nor ADAM17 correlate with age. * Refers to unspecific bands

Article Snippet: Representative blots of CSF samples of non-AD control individuals with large age amplitude (the age of the subjects are shown on top) probed with ( A ) anti-ADAM10 polyclonal antibody to ectodomain region (OAGA02442, Aviva) and ( B ) ADAM17 antibody towards the ectodomain region (AF9301, R&D Systems).

Techniques: Control

Journal: Alzheimer's Research & Therapy

Article Title: Selective reduction of ADAM10 in brain and cerebrospinal fluid of Alzheimer’s disease patients

doi: 10.1186/s13195-026-02007-6

Figure Lengend Snippet: Levels of ADAM10 and ADAM17 species in AD CSF samples. Analysis of CSF ADAM10 and ADAM17 in non-AD controls (NADC) and in AD patients. A Representative blot of CSF probed against anti-ADAM10 ectodomain antibody. Quantification of immunoreactive band values obtained from B iADAM10, C mADAM10 and D sADAM10. E Values of the ratio between mature vs immature species of ADAM10 (mADAM10/iADAM10). F Representative blot of CSF probed against ADAM17 and the quantifications of the immunoreactivity of the bands for G iADAM17, H mADAM17 and I sADAM17. J Result of the ratio mADAM10/iADAM10. The graphs represent mean ± SEM. Significant P < 0.05 values assayed by t-test are indicated. * Refers to unspecific bands

Article Snippet: Representative blots of CSF samples of non-AD control individuals with large age amplitude (the age of the subjects are shown on top) probed with ( A ) anti-ADAM10 polyclonal antibody to ectodomain region (OAGA02442, Aviva) and ( B ) ADAM17 antibody towards the ectodomain region (AF9301, R&D Systems).

Techniques:

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: ADAM10-mediated increased cleavage of CX3CL1 contributes to CI-promoted GC progression. ( A-C ) RT-qPCR and Western blot confirmed that CI increased the mRNA and protein expression of ADAM10 in tumor tissues( n = 3 per group). ( D ) Immunofluorescence assay suggested the increase of ADAM10 (Red) fluorescence intensity in tumor tissues. ( E-F ) Western blot confirmed that LPS increased the protein expression of ADAM10 in GC cell lines( n = 3 per group). ( G ) ELISA assay demonstrated that the ADAM10 antagonist GI254023X could attenuate the LPS-induced increase in soluble CX3CL1 levels in GC cells( n = 3 per group). ( H-N ) ADAM10 antagonist GI254023X inhibited LPS-promoted GC cells migration and invasion( n = 3 per group). ( O ) Schematic diagram shows the experimental protocol for determine the effect of ADAM10 inhibitor GI in suppressingsing CI-promoted tumor progression in mice( n = 5 per group). ( P-Q ) Left: Representative macroscopic images of tumors; Right: weight of tumors( n = 5 per group). ( R ) Concentration of CX3CL1 in the mice tumors detected by ELISA( n = 3 per group). Data are represented as mean ± SEM. For panel F, statistical significance was determined relative to the 0 µg/mL LPS group. For panels G , I , K , N , statistical significance was determined relative to the LPS group. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001(A, C: two-tailed unpaired Student’s t test; F , G , I , K , N , Q , R : one-way ANOVA with Dunnett’s multiple comparisons test).

Article Snippet: Blotting detection was performed using anti-ADAM10 antibody(Proteintech, 25900-1-AP) and anti-CX3CR1 antibody(Proteintech, 29819-1-AP), followed by incubation with appropriate HRP-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Immunofluorescence, Fluorescence, Enzyme-linked Immunosorbent Assay, Migration, Concentration Assay, Two Tailed Test

Journal: Scientific Reports

Article Title: Chronic inflammation promotes gastric cancer progression via ADAM10-mediated cleavage of CX3CL1

doi: 10.1038/s41598-026-39743-6

Figure Lengend Snippet: Graphical Abstract. CI induces the upregulation of ADAM10 expression in GC cells. Upregulated ADAM10 mediates the cleavage of membrane-bound CX3CL1 (mCX3CL1), thereby promoting the release of soluble CX3CL1 (sCX3CL1). The increased sCX3CL1 ultimately enhance GC cell proliferation and migration to accelerate tumor progression.

Article Snippet: Blotting detection was performed using anti-ADAM10 antibody(Proteintech, 25900-1-AP) and anti-CX3CR1 antibody(Proteintech, 29819-1-AP), followed by incubation with appropriate HRP-conjugated secondary antibodies for 1 h at room temperature.

Techniques: Expressing, Membrane, Migration

Journal: Materials Today Bio

Article Title: Cartilage organoids bridging bench to bedside: A steroid-free strategy for early osteoarthritis repair

doi: 10.1016/j.mtbio.2025.102688

Figure Lengend Snippet: Sino-organoids promoted chondrogenic differentiation and cartilage matrix formation. (A) Scratch assay and quantitative analysis demonstrated that sino-organoids promoted cell proliferation. (B) Live/Dead assay of BMSCs in different organoid groups on day 3. (C) Quantitative analysis of cell migration. (D) CCK-8 assay indicating comparable proliferation rates in different organoid groups. (E) Western blot of COL-II, ACAN, SOX9, ADAM10, and MMP13 in day 7 organoids. (F) Immunofluorescence results of chondrogenesis- and degradation-related proteins in organoids. Data are presented as mean ± standard deviation; ∗ P < 0.05, and ∗∗ P < 0.01.

Article Snippet: The blots were probed with primary antibodies against COL-II, ACAN, SOX9, ADAM10, and MMP13 (Cell Signaling Technology, USA), followed by horseradish peroxidase–conjugated secondary antibodies, and visualized by enhanced chemiluminescence.

Techniques: Wound Healing Assay, Live Dead Assay, Migration, CCK-8 Assay, Western Blot, Immunofluorescence, Standard Deviation

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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: ADAM10 (B-3) Mouse mAb , Santa Cruz Biotechnology , Cat# sc-28358; RRID: AB_626636.

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