(A) Western blot of whole cell lysates of JIR325 grown under normal conditions (TSB) or under iron-limited conditions (TSB containing 100 μM 2,2-dipyridyl, TSB + Dp). The cell lysates were probed with polyclonal antibodies to ChtD (anti-ChtD), ChtE (anti-ChtE) and Srt (anti-Srt). Bound antibodies were detected with HRP-conjugated goat anti-rabbit IgG (Millipore). The protein marker sizes (in kDa) are indicated on the left of the blot. (B) RT-PCR of chtD, chtE and srt on RNA isolated from JIR325 grown to mid-logarithmic phase under normal (TSB) and iron-depleted (TSB + 150 μM 2,2-dipyridyl, TSB + Dp) conditions. The chtD (576 bp), chtE (230 bp) and srt (276 bp) gene regions were PCR amplified using the appropriate primer pairs (S1 Table). DNA standard sizes (kb) are stated on the right of the gel. Lanes: RT- with reverse transcriptase (cDNA); NRT- no reverse transcriptase (RNA); NC- no template control (dH20); DNA- JIR325 genomic DNA; L-PCR markers (Promega).