Fig. 2

Validation of RNA-Seq data by qRT-PCR. Quantitative reverse transcription-PCR (qRT-PCR) using SYBR green assays was performed to validate gene expression levels obtained by RNA-Seq. Four genes [entS, fepC, enolase, colicin transporter (colicin-T)] were selected to validate the RNA-Seq results of WT, transconjugant and recipient. Two biological replicates and three technical replicates were used for each sample of two different conditions (ID and IR). Gene expression was normalized by gmk and adk reference genes. In the figure, each bar indicates the increased fold changes of that gene corresponding to the same strain in ID as compared to IR conditions. For example, in WT (SE163A) entS transcript was increased approximately 70-fold in ID as compared to IR. Error bars indicate standard error of mean (SEM) of gene expression of two biological replicates for each strain