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Fig. 3. LOS is required for cortactin tyrosine phosphorylation but not for ErbB2 or Src kinase activation. HBMECs (starved for 24 hours) were either not infected (–) or infected for 3 hours with the wild-type (WT) or isogenic rfaC ({Delta}rfaC), lgtA ({Delta}lgtA), lgtE ({Delta}lgtE) or mtrC ({Delta}mtrC) defective mutants of the 2C43 or ROU strains of N. meningitidis, as indicated. Inocula were adjusted to obtain similar adhesion events between wild-type and mutant bacteria. After lysis, ErbB2 receptor was immunoprecipitated and immunoblotted with an anti-phosphotyrosine antibody (PY) (A, top), Src was immunoprecipitated and subjected to an in-vitro kinase assay using acid-denatured enolase as a substrate (A, middle), or cortactin was immunoprecipitated and immunoblotted with an anti-phosphotyrosine antibody and the blot was reprobed with an anti-cortactin antibody to confirm that similar protein levels were immunoprecipitated (A, bottom, B). (C) Quantification by densitometry analysis (using NIH Image software) of cortactin phosphorylation induced by the wild-type and mutants of the 2C43 strain. Average values (± s.e.m.) are presented from four independent experiments.





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