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. 2011 Apr 21;7(4):e1002024. doi: 10.1371/journal.ppat.1002024

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Reynolds et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Overnight C. difficile cultures of WT, ΔrecV(pRecV+), ΔrecV(pRecV_Y176F), ΔrecV OFF (×4) and ΔrecV ON (×4) were grown and analysed in the following ways: A. S-layer extracts were prepared and analysed by SDS-PAGE and Coomassie staining. B. Western blot of S-layer extracts using anti-CwpVrptI antibody (top; 1∶5000) and anti-CwpVNter antibody (bottom; 1∶5000). ◂, repeat domain; ◃, N-terminal domain. C. Orientation-specific PCR amplifying the ON and OFF orientations of the cwpV switch. D. Immunofluorescence analysis of expression of CwpV using anti-CwpVrptI antibody. Phase and fluorescence images of one field of view are shown.