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Fig. 7. Effect of PI 3-kinase inhibitors on bacterial entry and on actin polymerization. (A) Control HeLa cells or cells pretreated with 100 nM wortmannin or 50 µM LY294002 were infected with C. caviae GPIC. Four hours after infection the cells were fixed and extracellular and intracellular bacteria were labelled as described in the Materials and Methods section. The number of surface-associated and intracellular bacteria were counted in control and treated cells (n=number of cells) and the efficiency of entry (intracellular/total cell-associated) was calculated. Results are expressed as the efficiency of entry relative to that in control cells. This experiment is representative of two. (B) Control HeLa cells (top row) or cells pretreated for 30 minutes with 50 µM LY294002 (bottom row) were infected with FITC-coupled C. caviae GPIC, centrifuged and incubated for 5 minutes at 37°C before fixation and actin labelling using Alexa-546 phalloidin.
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