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Fig. 1. EPEC-infection induces annexin 2-p11 accumulation at PtdIns(4,5)P2-enriched sites of actin pedestal formation. HeLa cells, transiently expressing the different tagged proteins indicated, were infected with EPEC. Three hours after infection, cells were fixed, permeabilized and analyzed for the distribution of the respective, ectopically expressed protein. Staining with Texas Red-phalloidin (Tx-phalloidin) revealed the characteristic reorganization of the actin cytoskeleton beneath micro-colonies of adhering bacteria. Annexin 2 (anx2) -GFP and YFP-p11 accumulate at sites of EPEC attachment. PHD-YFP localizes to the same actin-rich structures, indicating elevated PtdIns(4,5)P2 levels at sites of pedestal formation. Since PtdIns4P 5-kinase is also enriched, the rise in PtdIns(4,5)P2 is possibly caused by its enzymatic activity. Bars 10 µm.





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