Fig. 6. Effect of VAPA and VAPB on ER-vesicle budding. VSVG^ts045-Myc was trapped in the ER and ER-vesicle budding was reconstituted in vitro in perforated cells. Released VSVG-containing vesicles were collected by centrifugation and subjected to quantitative western immunoblot. The amount of VSVG^ts045 in vesicles released from semi-intact cells incubated at 4°C was subtracted from that in cells incubated at 32°C and the values were normalised as a percentage of the total cellular VSVG signal. Vehicle (DMSO) control (solid bars), FFAT (grey bars) and AAAT peptides (white bars) were added at 62.5 µM final concentration to the budding assay where indicated. The values represent mean ± s.e.m.