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Figure 2


Fig. 2. Behavior of YPT mutants in anterograde transport and in a Kar2p secretion assay. (A,B) An ER-Golgi transport assay. The arrival of [35S]gp{alpha}F-HDEL in the Golgi was monitored by precipitation with {alpha}-1,6-mannose antibodies and protein A-Sepharose. The amount of gp{alpha}F-HDEL precipitated by {alpha}-1,6-mannose antibodies was normalized to the amount of budded gp{alpha}F-HDEL from the ER. The average and standard deviation from at least three independent experiments are shown. The ER-Golgi transport assay was performed at (A) 23°C and (B) 30°C. (C) Mutants in YPT1 secrete Kar2p. Colony blot of different YPT mutants grown overnight at 30°C. Secreted Kar2p was detected by immunoblotting.