Research Paper Volume 13, Issue 1 pp 77—88

Fbxo7 and Pink1 play a reciprocal role in regulating their protein levels

Specific interaction of Bag2 with Fbxo7. (A) the bacterially expressed GST-Fbxo7 or its mutants was co-incubated with the bacterially expressed His-Bag2 for in vitro binding assay. The pulled-down proteins were analyzed by western blot using anti-Bag2 antibody for recombinant His-Bag2 and anti-GST antibody for GST and GST-Fbxo7. (B), the plasmids carrying Fbxo7 or its mutants were co-transfected with a plasmid expressing Bag2 into HEK293A cells. Fbxo7 proteins were IPed by anti-Flag antibody. The IPed proteins were subjected to western blotting. (C,D) the plasmid expressing Fbxo7 were transfected with the plasmid expressing Bag2 or Bag5 into HEK 293A. The target proteins were IPed by anti-Flag or anti-Myc antibody and the IPed proteins were detected by western blot.

Figure 4. Specific interaction of Bag2 with Fbxo7. (A) the bacterially expressed GST-Fbxo7 or its mutants was co-incubated with the bacterially expressed His-Bag2 for in vitro binding assay. The pulled-down proteins were analyzed by western blot using anti-Bag2 antibody for recombinant His-Bag2 and anti-GST antibody for GST and GST-Fbxo7. (B), the plasmids carrying Fbxo7 or its mutants were co-transfected with a plasmid expressing Bag2 into HEK293A cells. Fbxo7 proteins were IPed by anti-Flag antibody. The IPed proteins were subjected to western blotting. (C,D) the plasmid expressing Fbxo7 were transfected with the plasmid expressing Bag2 or Bag5 into HEK 293A. The target proteins were IPed by anti-Flag or anti-Myc antibody and the IPed proteins were detected by western blot.