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. 2014 Sep 25;3(6):836–848. doi: 10.1002/mbo3.199

Figure 7.

Figure 7

Kinase activity of LuxQ is required for SypK activation of qrr1 expression. (A) Levels of qrr1 expression in ΔluxQ strain KV6529 harboring a luxQ-FLAG variant plasmid (vector [pVSV105], luxQ [pVAR48], luxQ [A216P] K+/P− [pVAR50], or luxQ [H378A] K−/P+ [pVAR51]) and a sypK plasmid (vector [pKV282] or sypK++ [pVAR70]). The wild-type strain ES114 harboring pVSV105 and pKV282 was used as the nonfluorescent control for quantifying fluorescence levels. The copy of sypK in plasmid pVAR70 contains the FLAG tag. Graphical and error bars represent the averages and standard deviations of triplicate biological replicates, respectively. Experiment was performed three times, with similar results. (B) Western blot of ΔluxQ strain KV6529 harboring a luxQ-FLAG variant plasmid (WT [pVAR48], K+/P− [pVAR50], or K−/P+ [pVAR51]) and a sypK plasmid (vector [pKV282] or sypK++ [pVAR70]). Anti-FLAG antibodies were used to detect the variants of LuxQ-FLAG. Numbers to the left indicate molecular-weight marker positions (in kDa). A nonspecific band detected throughout the samples is designated as “n.s.”. A band corresponding to SypK-FLAG was not detected in this experiment. Experiment was performed twice, with similar results.