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. 2001 Oct 1;155(1):133–144. doi: 10.1083/jcb.200106148

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Copyright © 2001, The Rockefeller University Press

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Figure 6. — Localized adhesion of N. meningitidis leads to src association with phosphorylated ErbB2. HBMECs (starved for 18 h) were either uninfected (−) or infected with the wild-type (WT), or the PilT-defective mutant (PilT⁻) of the 2C43 strain of N. meningitidis for 4 h before lysis. As control, cells pretreated the same way were stimulated with 10 ng/ml EGF for 3 min before lysis. (A) ErbB2 was immunoprecipitated and immunoblotted with an anti-src antibody (top). Blot was reprobed with an anti-ErbB2 antibody (bottom) to show that similar protein levels were immunoprecipitated. (B and C) Whole cell extracts were incubated with 5 μg of a GST-tagged peptide containing the SH2 domain of src and coupled to glutathione-Sepharose. The complexes were immunoblotted with an antiphosphotyrosine antibody (4G10) (top). Blots were reprobed with an anti-ErbB2 antibody (middle) and anti-EGFR antibody (bottom). Three independent experiments were carried out with similar results and representative results from one experiment are shown.