Figure 6.

Association of human Gaa1p and Gpi8p shown by coimmunoprecipitation. Plasmids for FLAG- or GST-tagged proteins were transiently transfected into CHO cells in the indicated combinations. NP-40 extracts were prepared 2 d later and were first incubated for precipitation with anti-FLAG beads. The precipitates were analyzed by Western blotting with anti-FLAG (lanes 1–3) and anti-GST (lanes 4–6) antibodies. The supernatants of the first precipitation were then subjected to precipitation with glutathione beads and Western-blotted with anti-GST antibody (lanes 7–9). Double asterisks indicate the heavy chain of anti-FLAG antibody, and single asterisks indicate nonspecific bands of unknown origin (lanes 4–6). Molecular markers in kilodaltons are indicated on the left.