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. 2015 Jun 2;11(6):810. doi: 10.15252/msb.20145880

Figure 2.

Figure 2

Proteomic analysis of BCR signalosomes
  • A The Venn diagram shows the overlap between the proteins that were enriched in BCR signalosomes at 5 and 15 min.
  • B A network view of proteins present in BCR signalosome. Proteins are color-coded based on their association with BCR signalosomes at 5 or 15 min after BCR cross-linking.
  • C Validation of the dynamic association of proteins with BCR signalosomes. A20 cells were stimulated with biotinylated α-IgG F(ab′)2 for the indicated time points, the signalosomes were isolated by streptavidin pull-downs, and enrichment of the indicated proteins was analyzed by immunoblotting.
  • D, E ANKRD13A associates with BCR signalosomes. A20 cells were transiently transfected with GFP-tagged ANKRD13A WT and the ANRKD13A ΔUIM mutant (D), or ANKRD13A WT and ANKRD13A UIM3/4 mutant (E). BCR signalosomes were isolated as described in (C), and the enrichment of ANKRD13A was probed using α-GFP antibody. The expression of GFP-tagged ANKRD13A WT, ANRKD13A ΔUIM, and ANKRD13A UIM3/4 mutants in the input material was verified.