Dic chambers upon adding indicated concentration of Cm; error bars give 95 CI assuming a binomial distribution. Bar color indicates growth rates of expanding cells, with the relative growth price given by the scale bar on the ideal. (D) Development curves at distinct Cm concentrations, offered by the size of increasing colonies (y-axis) within the microfluidic device. The deduced development rates dropped abruptly from 0.35 hr-1 (green squares) at 0.9 mM Cm to zero at 1.0 mM Cm (black triangles). (E) As in panel C, but for immotile wild form cells (EQ4m) that showed no significant corROS Kinase Compound relation among growth rate and fraction of growing cells (s 0.1). (F) Fraction of Cat1 cells remaining right after the batch culture Amp-Cm enrichment assay (fig. S5). The results (fig. S7) reveal important fractions of non-growing cells nicely above the basalScience. Author manuscript; readily available in PMC 2014 June 16.Deris et al.Pagelevel of organic persisters ( 10-3), for [Cm] 0.four mM till the MIC of 1.0 mM above which no cells grew. Error bars estimate SD of CFU, assuming Poisson-distributed colony appearance.NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptScience. Author manuscript; accessible in PMC 2014 June 16.Deris et al.PageNIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author ManuscriptFigure three. Growth-mediated feedback(A) Elements of interactions defining the feedback model. Every single link describes a relation substantiated in panels (B)D) (clockwise). (B) The connection among the internal and external Cm concentration ([Cm]int and [Cm]ext respectively), described by the red line, is obtained by balancing the passive influx of Cm in to the cell (Jinflux, Eq. [1]) using the price of Cm modification by CAT (JCAT, Eq. [2]). This nonlinear relation is characterized by an (red approximate threshold-linear kind, with a “threshold” Cm concentration, arrow), under which [Cm]int is kept low because the capacity for clearance by CAT nicely exceeds the Cm influx; Eq. [S12]. For , CAT is saturated and Jinflux Vmax (dashed grey line). (C) The expression levels of constitutively expressed CAT (green) and LacZ (black) reporters (IDO1 Purity & Documentation reported here in units of activity per OD (42)) are proportional towards the development rate for growth with sub-inhibitory doses of Tc and Cm respectively. (D) The doubling time (blue circles) of wild kind (EQ4) cells grown in minimal medium with several concentrations of Cm increases linearly with [Cm] (Eq. [4] and Box 1). I50 (dashed vertical line) provides the Cm concentration at which cell growth is decreased by 50 . Here, [Cm]int [Cm]ext as a result of the absence of endogenous Cm efflux for wild kind cells in minimal media (41) (see also Eq. [S9]). Every single point represents a single experiment; error bars in the doubling instances are common error of inverse slope in linear regression of log(OD600) versus time.Science. Author manuscript; offered in PMC 2014 June 16.Deris et al.PageNIH-PA Author ManuscriptFigure 4. Development price predictions and phase diagram(A) Growth price of Cat1 strain in minimal medium batch culture with varying Cm (filled circles) agrees quantitatively together with the prediction on the development feedback model (line) according to the measured MIC (dashed red line). Error bars SD; n 3. Dashed blue line would be the theoretical MCC. Diamonds indicate drug levels at which enrichment experiments identified important numbers of non-growing cells (fig. S7). (B) The MCC (blue line) and MIC (red line) predicted by the growth feedback model for strains with di.
