Figure 3.

Expression and membrane localization of mouse EDEM in yeast cells. mEDEM-HA was cloned into high (p423TEF) and low-copy (p413TEF) yeast expression vectors and expressed in a Δ htm1 PDR1-3 yeast strain carrying human CFTR. Cells were grown to an A₆₀₀ of ∼1.2 and lysed with glass beads. Membranes were separated from supernatant and solubilized in buffer containing SDS and Triton X-100. Samples were separated by SDS-PAGE and blotted on nitrocellulose. Immunodetection was carried out with HA antibodies (Babco) and ECL (Amersham Biosciences UK). The empty vector p413 TEF was taken as a control. The protein was solely detectable in pellet fractions (P). The supernatant (S) was clear.