FIG. 6.
ADP-ribosylation regulates NFAT DNA binding. (A) Recombinant NFATc2 DBD was incubated with 32P-labeled NFAT DNA binding element in the presence and absence of purified PARP-1 proteins for gel mobility shift assays. Effect of NAD+ and PJ-34 on the formation of NFAT-DNA complex was also examined. The amounts of NFAT-DNA complexes were quantified by using a PhosphorImager. (B) Wild-type or Ala418 NFATc4 DBD expressed in COS cells was incubated with various amounts of 32P-labeled NFAT DNA binding element (100, 150, and 200 fmol) for gel mobility shift assays. Similar amounts of wild-type or Ala418 NFATc4 DBD in gel shift assays were also shown. +, present; −, absent.