Detachment under shear flow of the wild-type TIL448, the pilin mutant TIL1289 and the mucus-binding protein mutant TIL1290.

<p>Shear-flow induced detachment profiles of <i>L. lactis</i> bacterial cells attached to PGM-coated polystyrene in PBS; wild-type TIL448 (black square); pilin mutant TIL1289 (white square) and mucus-binding protein mutant TIL1290 (light grey square). The given results are the average values and standard deviations over at least three different coupons and independently grown cultures.</p

Adhesion forces and percentage of non-adhesive, non-specific adhesive and specific adhesive events obtained using AFM force spectroscopy in milliQ-grade water on TIL448, TIL1230, TIL1289 and TIL1290 lacto-probes in contact with PGM-coated polystyrene.

a<p>NA: non-adhesive event.</p>b<p>NSA: non-specific adhesive event.</p>c<p>SA: specific adhesive event.</p

AFM force spectroscopy for the wild-type TIL448 and the plasmid-cured derivative TIL1230.

<p>(A, D) Representative SEM images of <i>L. lactis</i> bacterial cells immobilized onto AFM tip and cantilever; (B, E) histograms of adhesion forces and (C, F) typical force-distance curves obtained when probing interactions between the wild-type TIL448 (A, B, C) and the plasmid-cured derivative TIL1230 (D, E, F) and PGM-coated polystyrene using AFM force spectroscopy in milliQ-grade water. One representative experiment (1024 force curves) is shown.</p

AFM blocking assays with PGM or O-glycan fractions (total, acidic, neutral) purified from PGM for <i>L. lactis</i> TIL448 lacto-probe in contact with PGM-coated polystyrene in milliQ-grade water.

<p>The given results are the average values and standard deviations over five independent PGM-coated coupons and lacto-probes. Before blocking, the percentage of non-adhesive events for <i>L. lactis</i> TIL448 was 2% (see in the Results section).</p>a<p>NA: non-adhesive events.</p

AFM blocking assays with free PGM for the wild-type TIL448.

<p>(A, B) Histograms of adhesion forces and (C, D) typical force-distance curves for wild-type TIL448 lacto-probe in interaction with PGM-coated polystyrene, using AFM force spectroscopy in milliQ-grade water, before (A, C) and after (B, D) incubation with free PGM. One representative experiment (1024 force curves) is shown.</p

Composition of the acidic fraction of O-glycans released from PGM.

<p>Monosaccharide composition of individual glycans was established by MALDI-TOF-MS based on the <i>m/z</i> values of monosaccharides.</p><p>Nature of monosaccharides (Gal, GlcNAc and Fuc) was based on GC composition analysis and on previous reports <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079850#pone.0079850-Karlsson1" target="_blank">[48]</a>. <i>m/z</i> values of sialylated glycans correspond to [M+Na]⁺ adducts whereas sulphated glycans exhibited [M+2Na-H]⁺ adducts.</p

Composition of the neutral fraction of O-glycans released from PGM. Monosaccharide composition of individual glycans was established by MALDI-TOF-MS based on the <i>m/z</i> values of monosaccharides.

<p>Nature of monosaccharides (Gal, GlcNAc and Fuc) was based on GC composition analysis and on previous reports <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079850#pone.0079850-Karlsson1" target="_blank">[48]</a>. <i>m/z</i> values of neutral glycans correspond to [M+Na]⁺ adducts.</p

Bacterial strains used in this study.

a<p>Micalis, INRA, Jouy-en-Josas, France.</p

τ_W5%, τ_W50% and τ_W90% values (Pa) obtained in the shear stress flow chamber for <i>L. lactis</i> bacterial cells attached to PGM coating in PBS.

<p>The given results are the average values and standard deviations over at least three different PGM-coated coupons and independently grown cultures.</p