Fig. 6
Verification of the functional targets of TFs by EMSA and qRT-PCR. The original sequence peaks show the TFs binding regions, and TFs binding regions (PCR-amplified from P. aeruginosa PAO1 genome) identified by ChIP-seq were mixed with an increasing amount of purified TFs protein for the EMSA assay. The expression of target genes was examined by RT-qPCR at the same time. PhoB directly regulated the transcription of PA4139 (a), xcpR (b), nosR (c), PA0123 (d) and PA4108 (e). FleQ directly regulated the expression of PA4139 (f), PA3520 (g), PA3662 (h) and arnB (i). AlgR directly regulated expression of phdA (j). ExsA directly regulated the expression of popN (k), PA3842 (l) and ccoN2 (m). GacA directly regulated the expression of cdpR (n), glnD (o), nasA (p), cbpD (q) and magD (r). All experiments were repeated at least three times. Two-tailed Student’s t-tests were used to examine the mean differences between the data groups. *P < 0.05, **P < 0.01 and ***P < 0.001. Error bars show standard deviations
