Figure 5.

Coimmunoprecipitation of hVps35, hVps29, hVps26, and SNX1. (A) COS7 cells were transiently transfected with expression vectors encoding myc-tagged Vps35, Vps26, Vps29, and SNX1 as indicated. Total cell extracts were prepared in the absence of detergent and then immunoabsorbed with the indicated polyclonal antibodies followed by protein A Sepharose. Immune complexes were collected, washed, and subjected to electrophoresis followed by immunoblotting with an anti-myc antibody. (B) Microsomal fraction prepared from rat liver was immunoabsorbed, and the samples treated as described in A, expect that the immunoblots were probed with antibodies to hVps35 and hVps26 to detect the amount of each endogenous protein that was immuno-isolated.