Journal: The Journal of Biological Chemistry

Article Title: Alix-mediated selective packaging of β-catenin into extracellular vesicles enhances their proangiogenic function

doi: 10.1016/j.jbc.2026.111254

Figure Lengend Snippet: Alix mediates the interaction with β-catenin . A , HeLa cells were transiently transfected with 3HA-Alix ( green ) and 3FLAG-β-catenin ( red ). After 48 h, the cells were fixed and stained with a rabbit anti-HA antibody, followed by incubation with a tetramethylrhodamine isothiocyanate (TRITC)-conjugated goat anti-rabbit secondary antibody and a mouse anti-FLAG antibody, followed by incubation with an isothiocyanate-conjugated goat anti-mouse secondary antibody. The bar represents 50 μm. B , HEK293T cells (4 × 10 6 ) were transfected with 3HA-Alix and 3FLAG-β-catenin, and coimmunoprecipitation was performed with the FLAG antibody after 48 h. Cell lysates and immunoprecipitates were analyzed via Western blotting with anti-FLAG and anti-HA antibodies. The full-length blots are presented in . Alix, apoptosis-linked gene 2–interacting protein X; HEK293T, human embryonic kidney 293T cell line.

Article Snippet: Anti-HA (1:100 dilution, 51064-2-AP), anti-FLAG (1:100 dilution, 66008-4-Ig), and anti-β-catenin (1:100 dilution, 51067-2-AP) primary antibodies, along with their corresponding secondary antibodies, were all purchased from Proteintech.

Techniques: Transfection, Staining, Incubation, Western Blot