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FBPase is not engulfed in vesicles under the inactivation conditions used (see MATERIALS AND METHODS). At the indicated times after shift to glucose medium, cells were spheroplasted, hypotonically lysed, and incubated with proteinase K (K) and proteinase K with Triton X-100 (KT). Samples were then immunoblotted and analyzed with antibodies against ER lumenal Kar2p, Ha, and FBPase. In A, gid2Δ (YTS1) cells and in B cells (YTS3) chromosomally expressing Ha-tagged Gid2p were used.
