Fig. 2. Effects of mice intestinal contents on CPE:

A] MIC effects on CPE processing. MIC were prepared from 3 different mice [2 female and 1 male]. To evaluate their effects on CPE, 30 ng of native CPE was or was not mixed with 1 μl of each MIC in PBS and incubated at 37 °C for 10 mins. After this incubation, samples were boiled, electrophoresed on a 12% acrylamide gel and then processed for CPE western blot analysis. The same amount of each MIC alone [no CPE] in PBS was similarly CPE western blotted to assure that the observed immunoreactivity was due to the presence of CPE. B] Head-to-head CPE western blot comparison of the effects of MIC, purified trypsin or chymotrypsin on CPE processing. MIC alone [no CPE] was also included as a specificity control. Before electrophoresis and CPE western blotting as described for panel A, MIC or MIC plus CPE samples, but not CPE plus trypsin or chymotrypsin samples, were boiled. C] Head-to-head Coomassie blue staining comparison of the effects of MIC, purified trypsin or chymotrypsin on CPE processing. Samples were prepared and electrophoresed as in panels A and B except that 5 μg of CPE and/or 2.5 μl of MIC were added to samples [when specified]. Also electrophoresed were equivalent concentrations of MIC, purified trypsin or purified chymotrypsin in the absence of CPE, for comparison. Samples with MIC were boiled but other samples were not. Panel A, B and C results are a representative result of three repetitions. Numbers at left of gels or blots indicate size of standard proteins [in kDa].