The IpaC Carboxyterminal Effector Domain Mediates Src-Dependent Actin Polymerization during Shigella Invasion of Epithelial Cells
Figure 5
The C351 insertion in the IpaC effector domain does not prevent T3S translocation.
Bacterial strains were grown to exponential phase, and T3S was induced by the addition of Congo Red in the culture medium. Bacterial lysates (Lysate) or supernatants (CRsupe) were analyzed by anti-IpaB and anti-IpaC Western blot analysis. HeLa cells were challenged with the various bacterial strains for 30 min at 37°C. Translocated T3S effectors were subjected to immuno-precipitation and analyzed by Western-blot analysis (Transloc, Materials and Methods) or by anti-actin Western-blot analysis (actin). The corresponding T3S effectors are indicated on the right. Cells challenged with: C1: ipaC/pC1; C1+lat: ipaC/pC1 after latrunculin treatment; mxiD: the non-invasive mxiD strain; C351: ipaC/pC351; SF126: a polar insertion upstream of the ipa locus that leads to decreased expression of Ipa proteins [31].