FIG 3.
Regulation of cmrRST by c-di-GMP occurs independently of cmr switch orientation. (A) Colony morphology of cmr-Δ3 OFF and cmr-Δ3 ON containing either vector, pPtet::dccA, or pPtet::EAL. Strains were grown on BHIS-agar with 20 ng/mL ATc to induce DGC and EAL gene expression. (B) qRT-PCR analysis of cmrS transcript abundance in cmr-Δ3 OFF and cmr-Δ3 ON strains carrying pPtet::dccA or vector control grown in BHIS broth with 20 ng/mL ATc. Data were analyzed using the ΔΔCT method with rpoC as the reference gene and normalization to cmr-Δ3 OFF with vector. Shown are means with standard error of six biological replicates from two independent experiments. *, P < 0.05; ****, P < 0.0001; one-way ANOVA with Tukey’s multiple-comparison posttest. (C) Representative images of ΔcmrR::SNAP carrying pPtet::dccA or vector control grown in BHIS broth with 20 ng/mL ATc. Samples were imaged at ×60 magnification by light and fluorescence microscopy following staining with SNAP-Cell TMR-Star substrate. Percentages of total cells that were fluorescence-positive were determined from at least three fields each from two biological replicates. Shown are means and standard deviations.