Figure 5
Direct protein-protein interactions between NPDC-1 and RXR. Bacterially expressed flag epitope tagged NPDC-1 [NPDC(F)] (40 μg) was incubated with glutathione-agarose bound GST/human RXRα (GST-RXR) fusion protein in the absence or presence of bacterially expressed human RARγ (RAR) (60 μg) or in the presence of 9-cis retinoic acid (9cRA) (10-6 M) as indicated. NPDC(F) was also incubated with glutathione-agarose bound GST. After extensive washing, the retained proteins were resolved by gel electrophoresis and analyzed by immunoblot with anti-flag M2 antibodies. One percent (0.4 μg) of the available NPDC(F) input was also electrophoresed and a unique immunoreactive band of approximately 45 kDa representing NPDC(F) is indicated by an arrow. The position of prestained molecular weight standards are shown.