Fig. 5.
Failure of C2-ceramide to inhibit ARF1 or RhoA association with the plasma membrane.
PMNs were treated with C2-ceramide and stimulated as indicated for Figure 4. Cells were probe sonicated and fractionated on 15% to 35% discontinuous sucrose gradients as described in “Materials and methods” to isolate plasma membrane. Equal amounts of protein were run on 15% SDS-PAGE, and Western blots were probed with antibody against ARF1 or RhoA. U indicates unstimulated; CB, cytochalasin B; F, FMLP, and C2, C2-ceramide. One representative experiment of 3.