FIGURE 7.

Cr-TRP16 enhances NF-κB DNA binding activity by reducing cysteine residues on p65. A, effect of wild type or C15S Cr-TRP16 on NF-κB activation. HeLa cells were transfected with 1 μg of pcDNA3.1 or pcDNA-p65 or pcDNA-p65C38S expression plasmid and co-transfected with indicated amounts of wild type and mutant Cr-TRP16 for 36 h, and then nuclear protein extracts were prepared using a nuclear extraction kit (Active Motif) according to the manufacturer's instructions. Nuclear extracts from HeLa cells were assayed for NF-κB p65 activation using the TransAM NF-κB p65 kit. B, HeLa cells were transfected with 1 μg of pcDNA3.1 or pcDNA-p65 or pcDNA-p65C38S expression plasmid. The nuclear extracts were subjected to Western blot analysis. The level of poly(ADP-ribose) polymerase served as loading control.