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Fig. 6. Accumulation of phosphotyrosine-containing proteins and cortactin at the sites of S. aureus invasion. (A) FAK(+) or FAK(–) fibroblasts were infected with FITC-labelled S. aureus for 1 hour. After fixation and permeabilization, samples were stained with monoclonal anti-phosphotyrosine (P-tyr) antibodies. Whereas S. aureus induced massive accumulation of tyrosine-phosphorylated proteins in FAK(+) cells (arrows), no bacteria-associated tyrosine phosphorylation was evident in FAK(–) cells (arrowheads). (B) 293 cells were transfected with RFP-cortactin and infected for 1 hour with FITC-labelled staphylococci. In contrast to S. carnosus (arrowheads), S. aureus induced accumulation of cortactin (arrows). Bars, 10 µm. (C) Local recruitment of cortactin-RFP (green line) to cell-associated bacteria (red line) was quantified by plotting the fluorescence intensity along the indicated red lines as detected in the FITC- or RFP-channels, respectively, against the distance.
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