16 research outputs found

    Histograms of persistence length <i>L</i><sub><i>p</i></sub> (nm).

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    <p>The positions are provided by fitting WLC model to force spectroscopy data on the Pil strain and its pili-displaying derivatives with 1.5 Ī¼M BSA or 15 Ī¼M BSA for the Pil p<i>srtA</i> strain. Data were fitted using the Gaussian function (solid line) and Gaussian multi-peak analysis (solid line and black star (*)).</p

    Experimental configuration of single-molecule force spectroscopy assays.

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    <p>Scheme illustrating the measurement configuration that was used during the experiments with no force applied (A) and under force (B) when the stage was moving. C is a close-up of B.). The pilus attached to the trapped bead is disproportionally long for reasons of depiction. The 10.5-Ī¼m mounting bead (MB) was immobilized on the coverslip while the 1-Ī¼m probe bead (PB) was trapped by optical tweezers (OT). A piliated bacterium was non-specifically attached to the MB and a pilus to the PB. When the coverslip was moved, and the trap kept in a fixed position, a force was directly exerted on the pilus. (C) Assuming adhesion to be non-specific, the most likely situation is that a portion of the pilus was attached (white subunits) and not solely the adhesion pilin (red subunit). Only a part of the pilus (black subunits) was thus subjected to the applied force.</p

    Transmission electron micrograph of Pil strain.

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    <p>(A) Pil strain bacterium (scale bar is 500 nm). A close-up of a pilus (black square) is shown in (B) with higher contrast and (C) image analysis of the contour.</p

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

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    <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

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

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    <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

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

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    <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