Figure 3.

Both PX and BAR domain of SNX5 interacted with the C-terminus of VAChT.

A: Schematic diagram of VAChT and its deletion constructs. B: HA-VAChT was transiently transfected in COS7 cells for 48 hours, followed by cell lysates incubated with glutathione-sepharose immobilized GST-SNX5 and detected with anti-VAChT antibody by Western blotting. C: TacWT and TacA were transiently transfected in COS7 cells for 48 hours, followed by cell lysates incubated with glutathione-sepharose immobilized GST-SNX5 and analyzed with anti-IL2Rα antibody by Western blotting. D: COS7 cells were transiently transfected with TacA for 48 hours. Cell lysates were incubated with Ni-NTA agarose immobilized His, His-SNX5-PX and His-SNX5-BAR recombinant proteins and detected with anti-IL2Rα antibody by Western blotting. TacA: a chimeric protein fused VAChT C-terminus with an unrelated protein, Tac.