Figure 3

TLR independent NFκB activation by B. pseudomallei is not dependent on T3SS3 effectors. HEK293T cells were cotransfected with pNFκB-SEAP and mammalian expression vectors encoding genes for BopA (A) BopC (B) and BopE (C) for 24 hr. Supernatants were collected for SEAP assay (left panels). Total RNA was isolated for measuring of expression of effector genes (right panels) by real-time PCR. D) Cells transfected with BopE plasmid were lysed and analysed by Western blot with anti-BopE antibody. SopE was used as a positive control. Asterisks * and ** indicate significant differences of p < 0.05 and p < 0.01 between empty vector and plasmid expressing T3SS effector gene respectively.