Fig. 3.
Reconstitution of ER-to-Golgi trafficking of ceramide in perforated cells.16) (A) The procedure for the reconstitution of ceramide trafficking in semi-intact CHO cells is represented schematically. (B) Reconstitution of ceramide transport from the ER to the Golgi site for SM synthesis in the semi-intact cell system. Perforated wild-type (WT) and LY-A cells pulsed with [3H]sphingosine were chased at 37 °C for 30 min in the transport reaction mixture with the indicated combination of perforated cells (40 μg protein) and cytosolic fraction (100 μg protein). For the cytosol-minus experiments (−), no cytosolic fraction was added to the reaction mixture. The data are expressed as a percentage of the mean value of the wild-type control, where wild-type perforated cells (40 μg protein) were chased in the transport reaction mixture, containing wild-type cytosol (100 μg protein).