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. 2015 Jun 23;290(32):19597–19613. doi: 10.1074/jbc.M115.655654

FIGURE 4.

FIGURE 4.

In vivo binding of EpaA domains. A, expression of different EpaA domains using a heterologous S. cerevisiae expression system. S. cerevisiae strain BY4741 carrying either of the plasmids B2445 (control), BHUM2157 (no A), BHUM1983 (Epa1A), BHUM1985 (Epa2A), BHUM2150 (Epa3A), BHUM2018 (Epa6A), BHUM1988 (Epa7A), BHUM2051 (Epa8A), BHUM2020 (Epa9A), BHUM2152 (Epa11A), BHUM2153 (Epa12A), BHUM1987 (Epa13A), BHUM2154 (Epa15A), BHUM2155 (Epa19A), BHUM2156 (Epa20A), BHUM1993 (Epa21A) or BHUM1988 (Epa23A) was grown to logarithmic phase before EpaA domains were visualized at the cell surface by immunofluorescence microscopy using anti-HA primary and Cy3-conjugated secondary antibodies. Specific signals were quantified, and the values obtained are shown in percentage relative to the strain expressing the carrier domain alone (no A) set to 100. Scale bar corresponds to 10 μm. B, epithelial cell adhesion conferred by 15 EpaA domains. Adhesion was determined using S. cerevisiae strains shown in A or a C. glabrata strain (triple-auxotrophic derivative of CBS138) after 2 h of incubation with a monolayer of Caco-2 cells. Adhesion of S. cerevisiae strains expressing no adhesin (control) or only the carrier domain (no A) is shown in black. For S. cerevisiae strains, EpaA-specific adhesion is indicated by the white part of bars that results from subtracting the value obtained by the strain expressing the carrier domain alone (no A) from the total adhesion. Error bars indicate standard deviation obtained by 10 independent experiments. The functional classification is shown at the bottom. DIC, differential interference contrast.