Journal:

Article Title: Extracellular Signal-Regulated Kinase Activates Topoisomerase II? through a Mechanism Independent of Phosphorylation

doi:

Figure Lengend Snippet: Coimmunoprecipitation of endogenous topoisomerase IIα and ERK2 from nuclear extracts. ERK2 was immunoprecipitated from nuclear extracts prepared from nocodazole-treated NIH 3T3 cells; this was followed by immunoblotting to visualize both ERK2 and coimmunoprecipitating topoisomerase (Topo) IIα. Lanes 3 and 4 show increasing amounts of immunoprecipitated ERK2 and coimmunoprecipitated topoisomerase IIα. A small amount of topoisomerase IIα nonspecifically bound to the protein A-(Prot. A)-Sepharose resin (lane 2). Ten percent of the extract volume used for the immunoprecipitations served as a loading control (lane 1).

Article Snippet: For coimmunoprecipitations of endogenous ERK2 and topoisomerase IIα, nuclear extracts from nocodazole-treated NIH 3T3 cells were incubated with 0, 0.2, or 2 μg of anti-ERK2 antibody (C-14; Santa Cruz Biotechnology) for 2 h on ice; this was followed by addition of 20 μl of protein A-Sepharose (Pharmacia) that had been pretreated with BSA at 0.5 mg/ml.

Techniques: Immunoprecipitation, Western Blot