A. HEK293 cells were co-transfected with plasmids expressing TRADD and Flag-tagged NPMRAR or empty vector. Lysates were incubated with anti-FLAG antibody, and complexes captured on Protein A sepharose beads. Complexes were separated on SDS-PAGE, and immunoblots probed with anti-TRADD and anti-RARA antibodies.
B. HEK293 cells were co-transfected with expression plasmids for NPMRAR and V5_His-TRADD or V5_His empty vector. His-TRADD or His-empty vector complexes were captured on Ni-agarose beads and separated on SDS-PAGE. Immunoblots were probed with antibodies against RARA, TRADD, and the C-terminus of NPM.