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. 2007 Jan 25;117(2):428–437. doi: 10.1172/JCI29625

Figure 5. Cell-surface expression of PAT-1.

Figure 5

Caco-2 monolayers grown on plastic supports were infected with nonpathogenic E. coli or EPEC for 60 minutes and subjected to biotinylation at 4°C using sulfo-NHS-biotin. Surface and intracellular fractions were run on 10% SDS-polyacrylamide electrophoresis gels, followed by transfer to nitrocellulose membrane. The blot was immunostained with an avidin-peroxidase antibody (A) or rabbit anti–PAT-1 (B). Representative blots of 3 different experiments are shown. (C) Scanning densitometry of the PAT-1 protein band was performed and expressed as surface/total PAT-1 (surface plus intracellular PAT-1). The intensities of EPEC and nonpathogenic E. coli were calculated in relation to those of the uninfected cells, and the value of each time control was arbitrarily set to 100. Values represent mean ± SEM of 3 different blots.