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Fig. 2. Influence of erv46 and vma21 deletion mutations on ${alpha}$ factor precursor transport and packaging into COPII vesicles. (A) Washed semi-intact wild-type (BY4742), erv46 ${Delta}$ (SOY01409), vma21 ${Delta}$ (SOY01629) or erv46 ${Delta}$ vma21 ${Delta}$ (SOY01655) cells containing ³⁵S-labelled gp ${alpha}$ f were incubated without (NA) or with purified COPII subunits, Uso1p and LMA1 (Recon) in the presence of an ATP regeneration system. After 90 minutes at 23°C, outer-chain modified ³⁵S-gp ${alpha}$ f was immunoprecipitated and amounts were plotted relative to ³⁵S-gp ${alpha}$ f precipitated on concanavalin A beads. (B) Washed microsomes prepared from the same strains as in A containing ³⁵S-gp ${alpha}$ f were incubated without (NA) or with purified COPII subunits (COPII) in the presence of an ATP regeneration system. After 20 minutes at 23°C, the relative amounts of gp ${alpha}$ f in the diffusible vesicle fraction were measured. All assays were performed in duplicate and error bars represent the range.

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