Fig. 6

Translocation of PKCι induced by PAF and its insensitivity to Ro 31-8220 treatment. HEC-1B cells were incubated in the absence of serum for 24 hr prior to stimulation with 1 nM PAF either alone (A) or in the presence of the PKC inhibitor, Ro 31-8220 (B) for the times indicated. Cells incubated with PAF in the presence of 10 µM Ro 31-8220 were pretreated for 30 min prior to stimulation. Cytosolic and particulate fractions were prepared and analyzed by Western immunoblot analysis with PKCι antibody. Top panel: Western blot showing location of the iota isozyme in resting confluent cells, its association with the membrane following PAF addition (A) and failure of Ro 31-8220 to inhibit this association (B). Bottom panels of (A) and (B): Corresponding densitometric analysis. Presented are the results of a typical experiment (n = 4).