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1.
Fig. 1.

Fig. 1. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Schematic representation of fusion proteins expressed on the surface of S. cerevisiae using Gateway expression vector pBC542. Specific CWP sequences from C. albicans (Als3p, Eap1p, Hwp1p, Rbt1p) or S. cerevisiae (Cwp1p) are fused to an HA-tagged construct with the Ser/Thr-rich region of C. glabrata protein Epa1p and the C-terminal domain of S. cerevisiae protein Cwp2p carrying a GPI anchor. Amino acid residues for each segment are indicated.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
2.
Fig. 5.

Fig. 5. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Attachment to silicone by S. cerevisiae cells expressing candidal adhesins. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were incubated with silicone squares for 4 h at 30°C. Nonadherent cells were removed, and total biomass was measured by crystal violet staining. Values given represent mean ± SD of results of two independent experiments performed in triplicate.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
3.
Fig. 2.

Fig. 2. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Confirmation of surface expression of C. albicans adhesins by surrogate host S. cerevisiae. C. albicans SC5314 adhesins Als3p (small [sm] and large [lg] alleles), Eap1p, Hwp1p, and Rbt1p were cloned into S. cerevisiae BY4742 together with S. cerevisiae control protein Cwp1p and pBC542 vector alone. Expression clones were then fixed in paraformaldehyde, probed with antibody directed against the HA tag incorporated within each fusion protein followed by a FITC-conjugated secondary antibody, and visualized by fluorescence microscopy (scale bar, 50 μm). Arrows indicate examples of cell clumping.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
4.
Fig. 8.

Fig. 8. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Effects of Congo red on the interactions of Streptococcus gordonii DL1 with S. cerevisiae expressing C. albicans adhesins. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were grown in CSM for 3 h in the presence of 30 μM Congo red and incubated for a further 1 h with FITC-labeled streptococci. Cells were then harvested and visualized by fluorescence and light microscopy (scale bar, 50 μm). Arrows indicate examples of streptococci attached to yeast cells.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
5.
Fig. 7.

Fig. 7. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Interactions of Streptococcus gordonii DL1 with C. albicans adhesins expressed on the surface of surrogate host S. cerevisiae. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were grown at 30°C in CSM for 3 h and incubated for a further 1 h with FITC-labeled streptococci. Cells were then harvested and visualized by fluorescence and light microscopy (scale bar, 50 μm). Black arrows indicate locations of streptococci in phase-contrast images. White arrows indicate examples of interactions between streptococci and aggregates of S. cerevisiae cells.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
6.
Fig. 4.

Fig. 4. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Attachment and biofilm formation on polystyrene by S. cerevisiae cells expressing candidal adhesins. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were incubated in polystyrene wells for 4 h at 30°C. Nonadherent cells were removed and total biomass was measured immediately by crystal violet staining (A) or after a further 20 h of incubation at 30°C in CSM (B). (C) Biofilm formation was visualized after 24 h by light microscopy. Values given represent mean ± SD of results of two independent experiments performed in triplicate.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
7.
Fig. 6.

Fig. 6. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Attachment and biofilm formation on extracellular matrix proteins by S. cerevisiae cells expressing candidal adhesins. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were incubated with 0.5 μg ECM protein for 4 h at 30°C. Nonadherent cells were removed, and total biomass was measured immediately by crystal violet staining (A) or after a further 20 h of incubation at 30°C in CSM (B). Values given represent mean ± SD of results of two independent experiments performed in triplicate.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
8.
Fig. 9.

Fig. 9. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Binding of Streptococcus gordonii DL1 to S. cerevisiae expressing C. albicans adhesins in the presence or absence of Congo red. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were grown in CSM for 3 h in the presence (open bars) or absence (filled bars) of 30 μM Congo red and incubated for a further 1 h with FITC-labeled streptococci. Cells were then harvested and visualized by fluorescence and light microscopy. Levels of streptococcal attachment were defined as the number of fluorescent pixels bound per Saccharomyces cell.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.
9.
Fig. 3.

Fig. 3. From: Heterologous Expression of Candida albicans Cell Wall-Associated Adhesins in Saccharomyces cerevisiae Reveals Differential Specificities in Adherence and Biofilm Formation and in Binding Oral Streptococcus gordonii.

Attachment and biofilm formation on a saliva-coated surface by S. cerevisiae cells expressing candidal adhesins. S. cerevisiae cells expressing candidal adhesins Als3p, Eap1p, Hwp1p, and Rbt1p, together with control protein Cwp1p and pBC542 vector alone, were incubated with saliva-coated coverslips for 4 h at 30°C. Nonadherent cells were removed, and total biomass was measured immediately by crystal violet staining (A) or after a further 20 h of incubation at 30°C in CSM (B). (C) Biofilm formation was visualized after 24 h by light microscopy (scale bar, 100 μm). Values given represent mean ± standard deviation (SD) of results of two independent experiments performed in triplicate.

Angela H. Nobbs, et al. Eukaryot Cell. 2010 Oct;9(10):1622-1634.

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