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[Preprint]. 2023 Jun 7:2023.06.06.543935. [Version 1] doi: 10.1101/2023.06.06.543935

Figure 2. Doxycycline-depletion of doxycycline-regulated ffmA transcription.

Figure 2.

A. A doxycycline-repressible (doxycycline off: DO) promoter-driven ffmA gene (DO-ffmA)-containing strain was grown overnight in the absence of doxycycline. A sample of this culture was processed to measure FfmA expression in the absence of doxycycline (0 hours) and then doxycycline was added for the numbers of hours indicated. Cultures were processed at each indicated time point and whole cell protein extracts prepared to determine the effect of doxycycline addition on expression of FfmA protein by western blotting with anti-FfmA. Tubulin was also analyzed as a loading control along with Ponceau S staining of the membranes after protein transfer. The numbers refer to relative expression normalized to the 0 hour time point. B. Samples from the strains grown as in A were processed for RNA and levels of either ffmA or act1 mRNA were determined by qPCR.