Relative expression of <i>wzz</i> mRNA (A and B) and LPS analysis (C) of <i>rcsB pmrA</i> double mutant.

<p>A,B. Total mRNA was harvested from cultures of SEΔ<i>rcsB</i>Δ<i>pmrA</i> double mutant and <i>S.</i> Enteritidis wild type #5694 grown in LB media (A) or grown in low Mg²⁺, low Mg²⁺+Fe³⁺ and high Mg²⁺ (B). The relative mRNA amount was determined by reverse transcription real-time quantitative PCR and related to mRNA levels in wild type strain (A) or in wild type strain grown in low Mg²⁺ (B), set as 1. Values are means ± SD of five independent mRNA extractions performed in triplicates. * significant difference p<0.01 with respect to wild type strain grown in the same media; † significant difference p<0.05 with respect to same strain grown in <i>pmrA-</i>inducing conditions (low Mg²⁺ and low Mg²⁺+Fe³⁺). C. Equal amount of LPS was loaded in each lane and analyzed by Tricine/SDS-PAGE on a 14% (w/v) acrylamide gel followed by silver staining. The concentration of LPS was determined by measuring KDO using the purpald assay.