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. 2013 Jan 4;2(1):161–172. doi: 10.1002/mbo3.54

Figure 2.

Figure 2

Reduced expression of exoT, exsD, and popN in crc deletion mutant. (A) and (B) Transcriptional expression of exsD (A) and exoT (B) in wild-type PAO1 and Δcrc mutant carrying a vector with PexsD–lacZ or PexoT–lacZ transcriptional reporter gene. (C) and (D) The translational expression of exsD (C) and popN (D) in wild-type PAO1 and Δcrc mutant carrying a vector with PexsD’-‘lacZ or PpopN’-‘lacZ translational fusion reporter genes. Bacteria were grown at 37°C in Luria–Bertani medium containing 5.0 mmol/L nitrilotiracetic acid. (E) Reduced cytotoxicity in crc mutant in comparison with wild-type PAO1. Cytotoxicity was assayed using Cell Counting Kit-8 (Dojindo Molecular Technologies), which is based on the dehydrogenase activity detection in viable cells. HeLa and A549 cells were infected with bacterial cells at multiplicity of infection of 50. Absorbance at 450 nm was measured and the cell viability was calculated. The data were the means of four (A–D) or five (E) replicates and error bar represents standard deviations.