Fig. 4

Zur is involved in CzcR expression. A) Growth curves of P. aeruginosa wt, Δzur, ΔczcRS and ΔcadR mutants in Zn-depleted medium (30 µM TPEN), in 2 mM and 4 mM ZnCl₂, as indicated. B) Expression of czcR measured using qRT-PCR. RNAs were extracted at 0, 15 min, 1 h and 5 h after addition of 2 mM ZnCl₂ as indicated. Fold expressions are compared to the wt strain at t0 and normalized by oprF expression. Standard deviations of the triplicates are indicated. Statistical analyses were performed according to the Student’s t-test and p-values are given as follows: ≤ 0.05 (*). C) Western blot analysis of total proteins sampled just before (0 h), or 1 h and 5 h after addition of 2 mM Zn of the wt and Δzur mutant, containing the empty plasmid (pE) or the IPTG inducible zur:6His gene (pzur). Blots were decorated with anti-CzcR, anti-6His for Zur detection and anti-Hsp70 as loading control