Fig. 2

QRT-PCR validation of selected genes in the virR and revR mutants. RNA was extracted from cells grown in TPG broth for 5 h, which corresponded to the late logarithmic growth phase. RNA was converted to cDNA using reverse transcriptase, and was subsequently used as the template in QRT-PCR, which was performed to validate the relative expression of the pfoA, ccp, plc, and cspB genes in the virR mutant (a) and the expression of the pfoA, plc, nirC and cspB genes in the revR mutant (b). Expression levels are shown relative to the expression of the housekeeping gene, rpoA, and are the average of at least three independent biological replicates (n ≥ 3, mean ± SEM). The asterisk (*) denotes a statistically significant difference (p ≤ 0.05) as calculated with the student’s unpaired t-test