Figure 3
From: Feedback inhibition by RALT controls signal output by the ErbB network
Mapping of the minimal EBR in the RALT protein. (a, top) Schematic outline of domain organization of RALT and RALT ΔEBR mutant. NLS indicates a putative nuclear localization signal. (Bottom) Schematic representation of RALT fragments used as GST fusion products in pull-down assays reported in panel b. The ability of each recombinant polypeptide to interact in vitro with the ErbB-2 protein is also indicated. Note that RALT fragments are drawn to a scale larger than the one used to draw RALT and RALT ΔEBR in the top portion of the panel. (b) GST fusion proteins (5 μg for each pull-down experiment) spanning the indicated portions of the RALT protein were produced in E. coli, purified onto glutathione–agarose beads and assayed for their ability to capture ErbB-2 protein solubilized from NIH-ErbB-2 transfectants. Protein complexes were analysed by immunoblot with anti-ErbB-2 antibodies. GST was used as control; reference lysate corresponded to 5% of input lysate in pull-down assays
