Figure 5.
ARF translocation to the plasma membrane in the reconstituted system was not inhibited by prior C2-ceramide treatment. Aliquots were taken from samples in experiments described in Figure 4. These aliquots were ultracentrifuged, and the pellets were solubilized in sample buffer to run 12.5% SDS-PAGE. Proteins were transferred, and Western blots were performed with antibody against ARF1. (A) A representative Western blot. U indicates unstimulated; G, GTPγS; C, C2-ceramide (5 and 10 μM); D, dihydro-C2-ceramide. (B) Scanning densitometry of the ARF1 band in Western blots. Data shown are the mean ± SEM of 5 experiments.
