FIG. 5.

Isolation of early endosomes and Western blotting revealed translocation of PKC β I after infection. J774 cells were infected with wild-type or mutant L. monocytogenes for 4 min, and purified endosome fractions were prepared. One hundred micrograms of protein of each sample (from two pooled endosome preparations) was used for SDS-polyacrylamide gel electrophoresis followed by Western blotting, as described in the text. PKC β I translocation was observed in cells infected with the wild type (lane 3) and the PI-PLC mutant (lane 6) compared to uninfected cells (lane 1). Translocation was not seen after infection with the LLO mutant (lane 2) or after 30 min of pretreatment with hispidin and infection with the wild type (lane 5). Cells pretreated with SK&F 96365 for 30 min showed translocation of PKC β I (lane 4).