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FIGURE 1. TFIIIA interacts with PIAS2b and XCtBP. A, colony lift filter assay used to measure interactions between PIAS2b and TFIIIA in a yeast two-hybrid system. Triplicate assays are presented in each column, with the following bait/prey combinations: lane 1, lamC/T antigen (negative control); lane 2, TFIIIA (fingers 1–3)/PIAS2b; lane 3, TFIIIA (fingers 1–4)/PIAS2b; lane 4, p53/PIAS2b. B, MBP-TFIIIA or MBP-L5 (negative control) fusion protein was incubated with reticulocyte lysate containing 35S-labeled PIAS2b. The fusion protein was recovered by binding to amylose resin. Precipitated samples were analyzed by SDS-PAGE followed by autoradiography. C, reticulocyte lysate containing 35S-labeled XCtBP was incubated with either TFIIIA or PIAS2b alone or with both proteins together. Complexes were collected by immunoprecipitation using either anti-TFIIIA (lanes 1 and 2) or anti-PIAS2b (lanes 3 and 4) antibody adsorbed to protein A-Sepharose. Control assays used unmodified protein A-Sepharose. Precipitated samples were analyzed by SDS-PAGE followed by autoradiography. Image published in: Malik MQ et al. (2014) Copyright © 2014. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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