758513.fig.005a
(a)
758513.fig.005b
(b)
Figure 5: Fibronectin and laminin degradation by plasmin-coated leptospires. Spirochetes were incubated in lsPBS with no additions (UT) and with addition of PLG and uPA (plasmin). ELISA plate wells coated with 0.5 μg/well fibronectin (a) or laminin (b) were incubated for 20 h with the spirochetes from each experimental group to allow substrate digestion. Plates were then incubated with antibodies antilaminin or antifibronectin. A reduction in absorbance was interpreted as substrate digestion, relative to controls lacking bacteria (0.5 μg/well purified laminin or cellular fibronectin, corresponding to 0% degradation). Percent degradation (a reduction in absorbance value relative to control lacking bacteria) was calculated as described in Section 2. Bars represent mean percent substrate degradation relative to the positive control (0% degradation) ± the standard deviation of three replicates. The experiment was independently performed three times with similar results. P < 0.001 (*) and P < 0.01(**).