JCS02328 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. S. aureus internalisation recruits vinculin to the sites of bacteria-host cell interaction. 293 cells were transfected with GFP-vinculin and infected with rhodamine-labeled S. aureus or S. carnosus at MOI 20 for 1 hour. Whereas S. aureus induced local accumulation of GFP-vinculin (small arrow), no vinculin recruitment was observed in the vicinity of cell-associated S. carnosus (arrowhead).

• Supplemental Figure 2 -

  Fig. S2. Host cell contact of S. carnosus does not lead to local accumulation of phosphotyrosine-containing proteins. FAK-re-expressing fibroblasts were infected with rhodamine-labeled S. aureus or S. carnosus at a MOI 20 for 1 hour. After fixation, samples were processed for immunofluorescence staining with monoclonal anti-phosphotyrosine antibodies and Cy2-conjugated secondary antibodies. In contrast to S. aureus (small arrows), S. carnosus did not induce local accumulation of phosphotyrosine-containing proteins at the site of cell contact.

• Supplemental Figure 3 -

  Fig. S3. Cortactin recruitment to S. aureus is not dependent on the presence of FAK. FAK(–) and FAK(+) cells were transfected with cortactin-RFP and infected with FITC-labelled S. aureus (Cowan) for 1 hour. The arrowheads point to cortactin accumulation at the site of cell-attached S. aureus. RFP-cortactin recruitment is evident in both FAK(+) and FAK(–) cells, suggesting that local accumulation of cortactin at bacterial attachment sites is independent of FAK. Bars, 10 µm.

• Supplemental Figure 4 -

  Fig. S4. Fibronectin-stimulated tyrosine phosphorylation of cortactin requires Src PTK activity. Serum-starved FAK(+) cells were seeded onto either poly-L-lysine (PL)- or fibronectin (FN)-coated dishes in the presence or absence of 5 µM PP2 for 45 min before cells were lysed. After cortactin immunoprecipitation (IP), samples were analysed by Western blotting with monoclonal anti-phosphotyrosine antibody (P.Tyr; upper panel). Following stripping, the blot was re-probed with polyclonal anti-cortactin antibodies (lower panel).