Figure 2.

RT-qPCR validation of RNA-sequencing (RNA-seq) data using six selected genes and wild-type, P. ananatis LMG 2665^T RNA samples. The fold change in gene expression levels between sampling points, before Quorum Sensing (QS) (wild-type strain at OD₆₀₀ = 0.2) and in the presence of QS (wild-type strain at OD₆₀₀ = 0.5) were calculated using the comparative cycle threshold (C_T) method [24] and data normalization was done using the ffh gene. The RT-qPCR results indicate fold change as cells shifted from before quorum sensing (OD₆₀₀ = 0.2) to during quorum sensing (OD₆₀₀ = 0.5) in LMG 2665^T. The fold change in RNA-seq data were calculated using 2^{(log₂FoldChange)}. Error bars represent the range of relative expression calculated using 2^{−(ΔΔC_T ± StandardDeviation)}. Triplicates were used per biological sample. The genes aroE and the one encoding for glycoside hydrolase were not differentially expressed in RNA-seq data (M5 versus W5).