Fig. 4.

Localization of NTAL interactors.A, isolation of nuclei (N), cytosol (C), detergent-resistant membrane (DRM), and detergent-soluble membrane (DSM) fractions from NB4 and U937 cells, prepared by differential detergent solubility followed by Western blotting (NTAL, LAMTOR1, LAMTOR5, Fyn/Yes, CD44, Lyn, SHIP1/INPP5D, and Hck). B, quality control Western blotting of specific markers for nonraft membranes, cytosol, and nuclear proteins. β-tubulin, appears in the cytosol fraction; LAMP1, lysosome-associated membrane protein 1, appears in the nonraft membranes; and fibrillarin is present only in the nuclear fraction. C, NTAL interactors were validated by counter-IP (co-IP) using antibodies against some of the identified proteins, followed by immunolabeling using NTAL antibody. IgG represents isotype negative controls for each experiment. IP, immunoprecipitation; NTAL, non–T cell activation linker.