Figure 1.

Reconstitution of the EE/RE-TGN transport by using a modified STxB. (A) Transport was carried out in the absence or presence of 0.6, 1.2, 1.8, 2.4, and 3.0 mg/ml rat liver cytosol as indicated. The relative level of transport was shown, with the highest transport level being defined arbitrarily as 100%. (B-D) Standard transport mixtures containing perforated cells, rat liver cytosol, and ATP regeneration system were preincubated for 1 h on ice with BFA (B), nocodazol (B), GTPγS (C), or various antibodies (D) as indicated. They were then shifted to 37°C and incubated for a further 90 min. Controls were the standard transport setups without any additional reagents or antibodies, and the levels of transport were arbitrarily defined as 100%. The respective concentrations (micrograms per milliliter) of the added reagents were shown in brackets (B and D). H, heat inactivated.