ev markers alix  (Cell Signaling Technology Inc)

Journal: Bioactive Materials

Article Title: ADGRG1-targeted hypoxia preconditioned extracellular vesicles ameliorate intervertebral disc degeneration by delivering taurine to disrupt the oxidative stress feedback loop-driven ferroptosis in nucleus pulposus cells

doi: 10.1016/j.bioactmat.2026.02.029

Figure Lengend Snippet: Preparation and characterization of engineered ADGRG1-targeting and hypoxia-treated EVs. (A)Induced fit docking analysis of ADGRG1-binding peptide (A1TP, 7 peptides) and extracellular domain of ADGRG1 protein (PDB database: 7SF8). (B) Analysis of the binding of the A1TP to purified ADGRG1 proteins using a microscale thermophoresis (MST) binding assay. (C) Induced fit docking analysis of A1TP-PEG and extracellular domain of ADGRG1 protein. (D) The binding free energy between A1TP or A1TP-PEG and ADGRG1 were calculated using molecular dynamics simulations. Lower values indicate more stable interactions, with values less than or equal to −20 considered as stable binding modes. (E) Schematic illustration of the conjugating reaction between DSPE-PEG-Alkyne and A1TP. Schematic illustration of the fabrication of A1TP-HX-EVs through external modification by A1TP anchoring. Specific steps for the synthesis of DSPE-PEG-A1TP (DPA) are shown in . (F) FT-IR analysis showed the characteristic peaks of the DSPE-PEG-A1TP. The new triazole ring itself showed a characteristic C=N stretching vibration, a peak at 1538 cm −1 revealed the successful conjugation of A1TP. (G) H Nuclear magnetic resonance (NMR) spectra of DSPE-PEG-A1TP in D2O. The hydrogen signatures of the phenyl and phenol groups at 7.5-8.0 ppm confirmed the successful conjugation of DSPE to A1TP. (H) Western blot analysis verified the presence of three EV marker proteins (ALIX, TSG101, and CD81) and one EV negative marker (GM130) in EVs, HX-EVs, and A1TP-HX-EVs. (I) Transmission electron microscopy (TEM) images of EVs, HX-EVs and A1TP-HX-EVs. Scale bar, 200 nm. (J) Zeta potentials of EVs, HX-EVs and A1TP-HX-EVs, n = 3. Two-tailed unpaired Student's t-test was used for statistical analysis. ns, not significant. A two-tailed unpaired Student's t-test was used for statistical analysis. (K) Representative images of the spherical morphology and dispersion states of EVs, HX-EVs and A1TP-HX-EVs. Scale bar, 500 nm. (L) Size distributions of EVs, HX-EVs and A1TP-HX-EVs.

Article Snippet: Finally, the presence of the characteristic EV markers Alix (92880, Cell Signaling Technology), CD81 (56039, Cell Signaling Technology) and TSG101 (sc-7964, Santa Cruz Biotechnology) was confirmed by Western blot analysis.

Techniques: Binding Assay, Purification, Microscale Thermophoresis, Modification, Conjugation Assay, Nuclear Magnetic Resonance, Western Blot, Marker, Transmission Assay, Electron Microscopy, Two Tailed Test, Dispersion

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

Article Title: A potential multimodal biomarker – cognitive signature associated with the conversion from subjective cognitive decline to mild cognitive impairment

doi: 10.1002/trc2.70240

Figure Lengend Snippet: Characterization and imaging of serum‐derived cEVs. (A) cEVs images as acquired by transmission electron microscopy; bar represents 100 nm. (B) Concentration and size distribution of cEVs examined by NTA using Nanosight‐NS300. (C) ApoA1 concentration in serum and cEVs. (D) Immunoblot detection of EV protein markers and non‐associated proteins. The parametric Student's t ‐test was used for statistical analysis of ApoA1; *** p < 0.001. ALIX, ALG‐2‐interacting protein X; ApoA1, apolipoprotein A1; CD63, cluster of differentiation 63; cEVs, circulating extracellular vesicles; CTR, controls; nm, nanometer; NTA, nanoparticle tracking analysis; SCD, subjective cognitive decline; TSG101, tumor susceptibility gene 101.

Article Snippet: Proteins were incubated for 30 min with primary antibodies against TSG101 (Novus Biologicals LLC, Catalog No.: NB200–112, 1:10), CD63 (R&D Systems; Catalog No.: MAB50482, 1:20), ALIX (R&D Systems; Catalog No.: MAB50482, 1:20), and calnexin (Novus Biologicals LLC; Catalog No.: NB100–1965, 1:25), followed by washing and incubation with species‐specific secondary antibodies.

Techniques: Imaging, Derivative Assay, Transmission Assay, Electron Microscopy, Concentration Assay, Western Blot

Journal: The Journal of Clinical Investigation

Article Title: Lysosomal homeostasis at the crossroads of neurodegeneration

doi: 10.1172/JCI199845

Figure Lengend Snippet: ( A ) The ESCRT machinery, recruited by galectin-3 (Gal3) and ALIX, polymerizes at rupture sites to reseal small pores. ( B ) The PI4K2A/ORP/ATG9A lipid-transfer axis forms ER-lysosome bridges that restore membrane lipid composition and curvature. ATG2 cooperates with ATG9A vesicles to deliver lipids for bilayer expansion. ( C ) Neutral sphingomyelinase–dependent conversion of sphingomyelin to ceramide promotes local fusion and sealing. ( D ) Under severe stress, protein-RNA condensates assemble into transient stress granule patches that shield lesions until structural repair completes. Together, these coordinated modules preserve lysosomal integrity and prevent cathepsin release.

Article Snippet: Galectin-3 interacts with the endosomal sorting complex required for transport (ESCRT) adaptor ALIX and TSG101 to nucleate ESCRT-III filaments that constrict and seal the membrane leak ( , ).

Techniques: Membrane