The effect of <i>Cj1411c</i> gene deletion on CPS production.

<p>Alcian blue stained CPS of <i>C. jejuni</i> 81-176 wild type (lane 2) was compared with the CPS extracted <i>C. jejuni</i> 81-176 Δ<i>Cj1411c</i> (lane 3) and <i>C. jejuni</i> 81-176 Δ<i>Cj1411c</i>::<i>Cj1411c</i> (lane 4).</p

CYP1411c spectral change following incubation with the inner membrane fraction.

<p>IMP o/n 1-5 represent successive spectral recordings following incubation of purified CYP1411c protein with <i>C. jejuni</i> 81-176 inner membrane proteins. Spectra were recorded at 1 min intervals.</p

CYP1411c cellular localization.

<p>(A) Anti-P450 immunoblotting on bacterial fractions (cytosolic fraction, Cyt; inner membrane protein, IMP and outer membrane protein fraction, OMP). Recombinant CYP1411c protein (purified) was used as a positive control. Fractions were validated with an antibody recognizing the cytosolic Fur protein (B) and the outer membrane protein CadF (C).</p

Virulence and serum resitance.

<p>(a) adhesion to HCT-8 cells, (b) invasion of HCT-8 cells of <i>C. jejuni</i> 81-176 wild type, <i>C. jejuni</i> 81-176 Δ<i>Cj1411c</i> and <i>C. jejuni</i> 81-176 Δ<i>Cj1411c</i>::<i>Cj1411c</i>. (c) serum resistance - the survival rate is defined as the number of <i>C. jejuni</i> 81-176 colonies isolated following exposure to human serum divided by the number of colonies surviving in heat inactivated serum, expressed as a percentage. Statistical significance (Student’s <i>t</i> test) relative to the level of wild type strain is indicated. *P=0.001. The experiments were done in triplicate and on three separate occasions. The error bars represent standard deviations for six separate wells.</p

qRT-PCR analysis of <i>Cj1411c</i> gene expression during infection.

<p><i>Cj1411c</i> expression analysis in <i>C. jejuni</i> 81-176 following infection of HCT-8 cells. The gene expression level for 1.5h in RPMI was set to 1 as the basal level. All other levels are expressed as fold change over the basal gene expression. Error bars represent ±S.D. of 3 independent experiments. The corresponding protein expression levels assessed by Western blot with the anti-P450 antibody are shown.</p