Known and putative virulence factor genes differentially expressed <i>in vivo i</i>n the <i>fliC</i> mutant compared to strain R20291.

<p>ND: non determined.</p

Flagellar genes differentially expressed <i>in vivo</i> in the <i>fliC</i> mutant.

<p>*From F1 region (late-stage flagellar genes) and **F3 region (early-stage flagellar genes) of the flagellar operon from <i>C. difficile</i> 630 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096876#pone.0096876-Aubry1" target="_blank">[39]</a>.</p

Functional clusters of <i>in vivo</i> differentially expressed genes.

<p>Differentially expressed genes in the <i>fliC</i> mutant compared to wild-type R20291 from caeca of 14 h post-infection mice were classified in functional groups according to their involvement in the biological process categories. (<b>A</b>) The number of analyzed genes is represented in green bars and the number of significantly differentially expressed genes is shown in black bars. Percentages of differentially regulated genes are indicated at right. (<b>B</b>) The numbers of up- and down-regulated genes for each cluster are indicated in green and black bars, respectively.</p

Membrane transport genes highly differentially expressed <i>in vivo</i> in the <i>fliC</i> mutant compared to strain R20291.

<p>Membrane transport genes highly differentially expressed <i>in vivo</i> in the <i>fliC</i> mutant compared to strain R20291.</p

Ability of sporulation of the <i>fliC</i> mutant compared to wild-type R20291 <i>in vivo</i> and <i>in vitro</i>.

<p>(<b>A</b>) Groups of 6 axenic mice were infected by oral gavage route with 1×10⁸<i>C. difficile</i> CFU. The <i>C. difficile</i> faecal vegetative cells and spores were measured by determining the concentration of CFU in faeces at 14 h post-infection by homogenising and plating on appropriated agar medium after heat shock (for spores) or not (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096876#s4" target="_blank">Materials and Methods</a>). Data represent the ratio of spores per vegetative cells. (<b>B</b>) Cultures in BHIS broth in anaerobic conditions were prepared from 2 successive subcultures as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096876#s4" target="_blank">Materials and Methods</a>. After heat shock, spores were quantified (CFU/ml) by performing serial dilutions and spread plating on BHIS agar supplemented with 0.1% bile salt taurocholate to induce germination. Data represent the ratio of spores per total cells. The presented values are the mean of 3 different cultures. Statistically significant difference is indicated by * for p<0,05.</p