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. 2008 Jan 10;27(2):447–457. doi: 10.1038/sj.emboj.7601976

Figure 6.

Figure 6

Kinetics of secretion of IpaC and truncated derivatives. (A) ipaC-mutant strains producing IpaC or IpaC derivatives were grown to exponential phase and incubated with Congo red to induce type III secretion (Materials and methods). Samples were centrifuged at the time points indicated (in minutes) above each lane. Bacterial pellets (top panels) and supernatants (bottom panels) were normalized to the total protein amounts and equivalent aliquots were analyzed by anti-IpaC western blot analysis (Materials and methods). (B) Quantification of the amounts of IpaC and IpaC derivatives in the bacterial pellets by scanning the band intensities in (A). Solid squares: IpaC; empty circles: IpaC Δ170–302; solid triangles: IpaC Δ101–302; empty squares: IpaC Δ299–363. The values are normalized to the initial values obtained at t=0, arbitrarily set up to 100. (C) Proteinase K digestion was performed on whole samples after induction of secretion for 30 min (Materials and methods). Bacterial pellets (P), supernatants (S), whole sample (W), and digested samples (D) were normalized and equivalent aliquots were analyzed by anti-IpaC western blotting.