Figure 5.

DNA protects TCP15 from inactivation by oxidants. TCP15 (770 ng of MBP-TCP15 fusion; 0.5 μm) purified with buffer lacking 2-ME was incubated in the presence of 10 mm diamide or 25 mm GSSG, as indicated, before or after the addition of DNA (100 ng; 0.25 μm double-stranded oligonucleotide labeled with 6-FAM). Controls (–) were incubated under similar conditions without redox agents. After incubation, the amount of TCP15-DNA complex present in each reaction was analyzed by EMSA. The amount of bound DNA relative to control lane 1 is specified above each lane and was calculated from signal intensities obtained with a Typhoon (GE Healthcare) scanner. The experiment was repeated at least three times with similar results.