Figure 5
From: Birth and Resuscitation of (p)ppGpp Induced Antibiotic Tolerant Persister Cells
Resuscitation of persisters is correlated with cessation of relBE transcription. MG1655valStsrpoS::mcherry cells carrying the pSEM4102 plasmid were grown in LB medium at 37 °C for about 6 hours (to mid-log phase) and treated with 100 µg/ml ampicillin for 3 hours, also at 37 °C. Survivor cells were concentrated by centrifugation and placed on a supplemented M9 agarose pad that contained β-lactamase to degrade any remaining ampicillin carried over from the growth medium. Changes in the cellular levels of YFPunstable and RpoS-mCherry were followed by fluorescence microscopy at 33 °C. Images were taken at the indicated time points. Combined mCherry (red) and YFP (green) channels are shown. Quantification of mCherry and YFPunstable fluorescence for 28 individual cells is shown in Fig. S5.
