Figure 7. NSP1 expressed in isolation binds to and induces degradation of β-TrCP.

293 cells were co-transfected with pFlag-β-TrCP and either pN-NSP1 or pO-NSP1. Forty hours post-transfection, MG132 was added to the media to a final concentration of 10 µM and the cells were incubated an additional eight hours. 10% of each sample was set aside as input and the remaining lysate was subjected to immunoprecipitation with anti-FLAG M2 Agarose Affinity Resin (Sigma). Immunoblots were probed with anti-Flag and anti-myc (NSP1 detection) antibodies.