Fig. 4: ADAP1 interacts with the immune signalosome.
a Representative immunofluorescence probing ADAP1 localization in unstimulated and stimulated (4 h TCR/CD28) TM. Repeated in 1 other donors with similar results. Scale bar is 10 μm for multicell images and 1 μm for single cell images. b Representative donor (n = 2) western blot analysis of cell fractionation assay. Resting (0 h) or stimulated TM (1 h, 4 h, 24 h) were fractioned into “S” supernatant (cytosolic) or “P” pellet (membrane) fraction. c Resting TM and stimulated (1 h) TM of one donor were immunoprecipitated in triplicate with ADAP1 antibody and submitted for tandem mass spectrometry analysis. Volcano plot represents average Log2Fc of factor enrichment in stimulated TM over resting TM. Samples with Log2Fc > 0.5 are highlighted in blue. A sample of immune signalosome proteins are highlighted. See Supplementary Data 1 for complete list of interacting partners, semi-quantitative abundance and statistical significance. d Visual representation of second donor mass spectrometry results of TM stimulated for 1 h and immunoprecipitated with IgG or ADAP1 antibodies. See Supplementary Data 2 for complete list of interacting partners, semi-quantitative abundance. Interacting proteins that form part of the immune signalosome or T cell membrane are highlighted in blue. e Protein–protein interacting network analysis clustered by gene ontology. Nodes are color-coded and size-coded based on p value. Edges (purple lines) represent network connections. Gene ontology category derived from Metascape analysis are written next to clusters. f Representative western blot analysis of resting or stimulated TM immunoprecipitated with ADAP1 or IgG antibodies (n = 2 donors). Samples were used for tandem mass spectrometry analysis (d, e). Elutions were additionally probed with PKCθ to validate mass spectrometry results. Source data are provided as a Source Data file.
