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. 2013 Mar 26;21(5):1076–1086. doi: 10.1038/mt.2013.41

Figure 1.

Figure 1

An overview of the P. aeruginosa “killed but metabolically active” T3SS-based vaccination. (a) The P. aeruginosa strain is cultivated under T3SS-inducing conditions. The protein used for vaccination is overexpressed by bacteria before bacterial inactivation with PCT in the presence of amotosalen. Then, P. aeruginosa is injected subcutaneously (s.c.), and its active T3SS allows protein injection into the host cells. (b) Colony-forming units (CFU) of OST and OSTAB after treatment with increasing amotosalen concentration. The mean of five experiments (performed in duplicate) is represented. (c) Analysis of the membrane integrity of the P. aeruginosa OSTAB strain with the Live/Dead BacLight bacterial viability kit (Invitrogen). OSTAB (live), heat-killed (HK), or photochemically treated P. aeruginosa in presence of 0–15 µmol/l amotosalen concentrations. (d) Western blot analysis of the protein S54-OVA in the supernatant of cultivated OSTAB S54-OVA strain with or without EGTA (i.e., closed or opened syringe), after PCT and in the presence of various amotosalen concentrations. IPTG was added for 3 hours after PCT. Representative blot of three experiments. EGTA, ethylene glycol tetraacetic acid; IPTG, isopropyl β-D-1-thiogalactopyranoside; PCT, photochemical treatment.