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. 2004 Jun 1;101(24):8870–8875. doi: 10.1073/pnas.0308605101

Fig. 5.

Fig. 5.

PIASy interacts with GATA-1 and specifically represses its transcriptional activity. (A) PIASy does not inhibit a GATA-1-independent promoter. Constructs expressing luciferase under control of human metallothionein minimal promoter (hMT), human c-myc promoter, or the GATA-responsive promoter was transfected in U2OS cells with or without T7-PIASy as described in Fig. 4. Experiments with the GATA reporter also contained expression vectors for the indicated GATA-1 proteins. Relative luciferase activity is expressed as percent of the activity of the promoters in the absence of PIASy. Lower shows the expression levels of PIASy in one representative experiment. (B) Interaction between PIASy and GATA-1. WT GATA-1 or the K137R mutant was transfected in 293T cells together with T7-PIASy as indicated. After 36 h, lysates were immunoprecipitated with a monoclonal antibody to GATA-1 and immunoblotted with an antibody to the T7 epitope (Right). Expression of GATA-1 and T7-PIASy was also analyzed in total lysates (Left). IgG heavy chains are indicated by an asterisk.