(A) HeLa cells (3–4 × 107) were transiently transfected with pCMV-myc-UPF1R(WT) or pCMV-myc-UPF1R(G495R/G497E) (10 µg). Two days later, total-cell lysate was immunoprecipitated using anti(α)-myc or mouse (m)IgG. Western blotting (WB) was performed using the specified antibodies. The four leftmost lanes analyzed decreasing amounts of protein before (−) IP.
(B) Essentially as in (A), except cells were transiently transfected with pCMV-myc-UPF1R(WT) and treated with the specified amount of OA for 3 hr prior to harvesting. All results are representative of three independently performed experiments.