Figure 2. Temporal FC analysis of P_iolR activity in MM with MI.

(A) MvP101 P_iolR::gfp or (B) MvP101 iolR::gfp grown in LB medium was adjusted to an OD₆₀₀ of 0.8, diluted 1:500 into MM/MI and incubated at 37 °C without agitation. At the indicated time points during growth, samples were collected and GFP-expressing cells were quantified by FC. The abscissa of each histogram represents the green fluorescence intensity at 515–545 nm on a bi-exponential scale, and the ordinate represents the numbers of bacteria relative to the maximal cell counts. Each histogram shows one representative data set of three biologically independent measurements. The inset illustrates the growth phase of the population during sample acquisition. (C) FC measurement of MvP101 P_rpsM::gfp as a control. The percentages of gfp-expressing cells (left) and non-fluorescent cells (right) are indicated. Samples were collected during a 48-h growth period after inoculation, and the average values and standard deviations of three independent growth experiments are shown; ***p < 0.001.