Fig. 4: Non-replicating persisters show equal metabolic activity compared to replicating STM WT in the early phase of infection. | Communications Biology

Fig. 4: Non-replicating persisters show equal metabolic activity compared to replicating STM WT in the early phase of infection.

From: The protected physiological state of intracellular Salmonella enterica persisters reduces host cell-imposed stress

Fig. 4

STM WT harboring double-inducible dual-fluorescence reporter was grown o/n in LB medium in the presence of arabinose. Before infection, arabinose was removed. RAW264.7 macrophages were infected, AHT was added 22 h p.i., and cells were lysed 24 h p.i. Liberated STM were recovered, fixed and subjected to FC. A Plotting the entire intracellular bacterial population against their arabinose-induced DsRed and AHT-induced sfGFP intensity shows distinct populations. As control, STM without addition of arabinose or AHT was analyzed. B The proportion of metabolically active and inactive NR STM WT at 22 h p.i. is shown. C Comparison of the metabolic activity of active NR and R STM WT 22 h p.i. D–H STM WT was cultured, infected and prepared for FC as described above. AHT was added at various time points p.i. and cells were lysed 24 h p.i. D FP levels anticipated for metabolically active and stress signal-positive NR STM. E Detected metabolically inactive NR STM may be stress signal-positive because of metabolic activity before addition of AHT. F Representative histograms of the metabolic activity of intracellular NR STM WT at various time points p.i. Arrows indicate increase in metabolically inactive NR STM, and decrease in metabolically active NR STM over time of infection. G Yellow, green, and magenta gates define subpopulations compared in H and I. H The proportion of metabolically active and inactive NR STM at the various time points was calculated. I Comparison of the metabolic activity of active NR and R STM WT at various time points p.i. J Green and orange gates define subpopulations compared in K. K Comparison of the metabolic activity of all NR and R STM WT at various time points p.i. L The ratio of metabolic activity of all NR STM to metabolic activity of R STM is shown. Means and standard deviations of one representative experiment are shown. Statistical analysis was performed as described for Fig. 2.

Back to article page