Figure 5.

SMN(K0) is capable of forming the SMN complex. (A) Empty pRK7 vector (mock), FLAG-SMN or FLAG-SMN(K0) was transfected in 293T cells. Whole-cell lysates were applied for immunoprecipitation using an anti-FLAG antibody, followed by immunoblotting of SMN. Asterisk denotes endogenous SMN (also applies for C). (B) An shRNA construct that targets GFP (negative control) or the 3′ UTR of SMN was used to establish stable SMN knockdown cells in 293T cells. Whole-cell lysates were used for immunoblotting of SMN and β-tubulin. (C) FLAG-GFP (negative control), FLAG-SMN or FLAG-SMN(K0) was stably expressed in 293T cells where endogenous SMN was stably knocked down shown in (B). Whole-cell lysates were applied for immunoblotting using indicated antibodies. (D) Using cells shown in (C), proteins co-immunoprecipitated with FLAG-SMN(K0) were immunoblotted using indicated antibodies.