FIGURE 5.

Gon4l, YY1, Sin3a, and HDAC1 can associate as parts of a single complex. 293T cells were co-transfected with the combination of expression plasmids indicated at the bottom of each panel. WCLs from transfected cells were subjected to sequential IPs. Nonspecific IgG was used to perform control IPs. Aliquots of material recovered from each IP were immunoblotted with antibodies that detect FLAG-Gon4l, Myc-Sin3a, YY1, or HA-HDAC1. The wedges at the top of the blots signify that serially increased amounts of WCLs were loaded. A, Gon4l, YY1, and Sin3a are all components of a single complex. Left panel, immunoblot analysis of proteins recovered by the first IP, which was performed using anti-FLAG antibodies. Right panel, immunoblot analysis of proteins recovered by the second IP, which was performed using anti-Sin3a antibodies. B, Gon4l, YY1, Sin3a, and HDAC1 are all components of a single complex. Left panel, immunoblot analysis of proteins recovered by the first IP, which was performed using anti-FLAG antibodies. Right panel, immunoblot analysis of proteins recovered by the second IP, which was performed using anti-HDAC1 antibodies. Immunoblotting for HA-HDAC1 was done using membranes already bound by YY1 antibodies, producing the signals marked with asterisks. All of the data shown are representative of at least three independent experiments. MW, molecular mass.