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. 2013 Mar 21;9(3):e1003361. doi: 10.1371/journal.pgen.1003361

Figure 3. AST2 interacts with PlRACK1.

Figure 3

A) PlRACK1 was identified as an AST2 binding protein using a far western assay with recombinant GST-AST2 or GST as a control. A HPT protein extract was subjected to SDS-PAGE, electroblotted to a PVDF membrane and overlayed with either GST (left) or GST-AST2 (right) alone and binding was detected using a GST antibody. B) The binding between recombinant AST2 with PlRACK1 was confirmed by a GST pull-down assay using GST-AST2 as the binding protein and proteins in a HPT cell lysate as bait. GST was used as a control in both assays. C) GST pull-down of His-Trx-AST2 by GST-PlRACK1. The bound proteins were analyzed by 12.5% SDS-PAGE. Bands corresponding to GST and GST-PlRACK1 were detected with an anti-GST antibody (lanes 1–4). The eluted material was also examined for the presence of AST2 with an anti-His antibody (lanes 5–8). Lanes 9 and 10 contain purified His-Trx-AST2 and HisTrx, respectively.