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. 2009 Mar 6;5(3):e1000325. doi: 10.1371/journal.ppat.1000325

Figure 7. Low CyaA or ET concentrations impair TCR–dependent Akt1 phosphorylation while enhancing Vav1 and p38 phosphorylation.

Figure 7

(A) Immunoblot analysis, using a phosphospecific antibody, of Akt1 activation in postnuclear supernatants from PBL activated for 5 min by CD3 cross-linking in the presence or absence of 45 nM CyaA or 110 nM ET (CyaA hi, ET hi) or alternatively in the presence or absence of 0.28 nM CyaA or 0.11 nM ET (CyaA lo, ET lo). A representative experiment is shown (n = 3). (B) Immunoblot analysis, using phosphospecific antibodies, of Vav1 (top) or p38 (bottom) activation in postnuclear supernatants from PBL activated for 1 min (Vav) or 5 min (p38) by CD3 cross-linking in the presence or absence of 45 nM CyaA or 110 nM ET (CyaA hi, ET hi) or alternatively in the presence or absence of 0.28 nM CyaA or 0.11 nM ET (CyaA lo, ET lo). Filters were stripped and re-probed with control antibodies. Representative experiments are shown (n≥4). The migration of molecular mass markers is indicated. The graphs on the right of the immunoblots show the quantification by laser densitometry of the relative levels of Akt1, Vav1, and p38 phosphorylation (phosphorylation in anti-CD3 stimulated cells taken as 100%) in PBL activated by CD3 cross-linking in the presence or absence of 45 nM CyaA or 110 nM ET (CyaA hi, ET hi) or alternatively in the presence or absence of 0.28 nM CyaA or 0.11 nM ET (CyaA lo, ET lo). ***P≤0.001**P≤0.01; *P≤0.05. Error bars, SD.