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. Author manuscript; available in PMC: 2012 Jul 13.
Published in final edited form as: Biol Chem. 2010 Jun;391(6):705–716. doi: 10.1515/BC.2010.066

Figure 1. Effect of Pseudomonas aeruginosa (PAO1)-conditioned medium on the integrity of elastase inhibitors elafin, SLPI and AAT.

Figure 1

8.3×10−7 M of elastase inhibitors (elafin, SLPI and AAT) were incubated with 10 μL PAO1- conditioned medium at 37°C and analysed by Western blot. (A) Time-course incubation of recombinant elafin with PAO1-conditioned medium for 0, 1, 2, 6 and 24h. Incubation products were analyzed by Western blot under reducing (upper panel) and non-reducing (lower panel) conditions. Arrow heads indicates cleavage products of elafin. (B) Identification of the Pseudomonas proteases involved in elafin cleavage. PAO1-conditioned medium was pre-incubated with the indicated proteases inhibitors (right panels) and mixed with recombinant elafin for 2h (upper panels) and 24h (lower panels). Elafin alone, PAO1-conditioned medium incubated in the absence (Con) or presence of elafin (+Elafin) are indicated in the left panels. Effect of PAO1-conditioned medium on (C) SLPI and (D) AAT integrity, and identification of the Pseudomonas proteases involved in their cleavage. PAO1-conditioned medium was pre-incubated with the indicated proteases inhibitors (right panels) before incubation with recombinant elafin for 24h. SLPI or AAT alone, PAO1-conditioned medium incubated in the absence (Con) or presence of SLPI (+SLPI) or AAT (+AAT) are indicated in the left panels.