A Novel Insight into Dehydroleucodine Mediated Attenuation of Pseudomonas aeruginosa Virulence Mechanism

Figure 5

Effect of DhL on Type III secretion. Top panel represents Type III secretion profile of P. aeruginosa strain PAO1 in presence of DhL as observed with SDS-PAGE stained with Coomassie blue stain. Type III null mutant (PA103 ΔUT) was used as a control. Lane 1 represents untreated PAO1 secretion profile and lane 2 represents treated PAO1 secretion profile. Under Coomassie blue stain, protein bands were observed at ~53 kDa, ~49 kDa, and ~37 kDa, indicating ExoT, ExoS, and ExoY, respectively, in PAO1 Type III secretion assay, which were not shown in DhL treated PAO1 Type III secretion assay. Lanes 3 and 4 showing untreated and DhL treated control strain (PA103 ΔUΔT) with no visible Type III secretion profile. Bottom panel represents immunoblot results of DhL treated PA103ΔUΔT: HA-ExoS strain carrying HA-tagged ExoS as described in Section 2. HA specific antibody was used in immunoblot (IB) to confirm the presence of ~49 KDa ExoS secretion. Experiments were repeated three times to confirm the observation.