Figure 5. Ndk deficiency leads to up-regulated expression of P. aeruginosa T3SS.
The indicated bacteria were inoculated into LB broth and a T3SS inducing condition (LB broth supplemented with 5 mM EGTA and 20 mM MgCl2) (initial OD600 of 0.01) and cultured at 37 °C for the indicated time. The bacteria and culture media were harvested for the subsequent analysis. (a) Real-time PCR analysis for T3SS-encoding gene expression. (b) Western blot analysis for the intracellular expression and secretion of T3SS proteins. (c) Real-time PCR analysis for T3SS gene expression with EGTA induction for 12 h. (d) Western blot analysis for intracellular T3SS protein expression with EGTA induction for 12 h. Representative images from triplicate experiments are shown (b,d). Data represent mean ± SD from three independent experiments (a,c). *Indicates P < 0.05 compared to PAO1 strain. #Indicates P < 0.05 compared to Δndk strain. ★ Indicates P < 0.05 compared to PAO1 + EGTA group. Δ Indicates P < 0.05 compared to Δndk + EGTA group.