Figure 3

Western Blots and co-immunoprecipitation of the SSG-2/SsPAQR1 interaction. Whole cell free extracts of S. cerevisiae cells containing pGBKT7 and pGADT7 plasmids with the complete SSG-2 coding region fused to the GAL4 activation domain and cMyc, and the initial insert coding fragment identified in the yeast two-hybrid assay fused to the GAL4 DNA binding domain and HA, respectively, were co-immunoprecipitated as described in Methods. The co-precipitated proteins were separated using 10% SDS polyacrylamide electrophoresis and transferred to nitrocellulose. The nitrocellulose strips were probed with anti-cMyc antibodies (Lane 1) and anti HA antibodies (Lane 3), respectively. Lanes 2 and 4 are negative controls where no primary antibody was added. The antigen-antibody reactions were detected using the Immun-Star™ AP chemiluminescent protein detection system. Pre-stained molecular weight markers were included in outside lanes of the gel and transferred to nitrocellulose, the position of the molecular weight markers is indicated in the figure.