Fig. 5. Effects of C3 exoenzyme on LTD₄-induced translocation of PKC, PKC and PKC to the membrane fraction. Cells were or were not pretreated with 5 µg/ml C3 exoenzyme together with 5 µg/ml lipofectamine for 10 hours and then were or were not stimulated with 40 nM LTD₄ (30, 60, 90 and 300 minutes). Thereafter, the cells were lysed and centrifuged at 200,000 g for 30 minutes as described in the Materials and Methods. The resulting membrane-rich pellet was separated by SDS-PAGE and immunoblotted with an anti-PKC, anti-PKC or anti-PKC antibody. The illustrated blots are representative of at least three separate experiments.