Figure 7.

GST-tagged TFI1, TFI2, and TFI3 interact with both Sed5p and COPI proteins. GST-tagged proteins were expressed in the cells of appropriate gene deletion strains, in which the GST-chimera was the only source of TFI1, TFI2, or TFI3 proteins. A membrane fraction was obtained after centrifugation at 150,000 g for 1 h at 4°C. Extracted membrane proteins (2 mg) were incubated with 50 μl prewashed glutathione–Sepharose beads. Each eluate was loaded on the 10% SDS-PAGE and analyzed by immunoblot with α-Sed5p and α-coatomer sera. The Coomassie blue staining of the samples presents on the left panel of the Fig. 1 B.