Fig. 4. Intracellular persister formation is not triggered by amino acid limitation nor ATP depletion.

a Intrabacterial ATP concentration in intracellular persisters (exposed to 50× MIC oxacillin for 24 h) and control samples (extracellular bacteria mixed with J774 cells lysate). Appropriate controls were performed to ensure the absence of contamination by eukaryotic ATP. Data are means ± SEM of three independent experiments. Statistical significance was determined by two-tailed Student’s t-test. ns non-statistically significant. b Schematic pathway of genes related to central carbon metabolism, annotated according to KEGG orthology⁷⁸ and Genbank database²³ and their log₂ Fold Change expression levels. c MA-plot of genes related to amino acid metabolism within the stringent response stimulon (Supplementary Fig. 4; see reference⁵⁵). The graph displays the log₂ Fold Change expression as a function of log₂ Base Mean (mean expression signal across all samples). Genes prominently activated after amino acid depletion²⁴ are pointed. The dotted line indicates the basal expression level in control samples. Statistical significance is based on adjusted P-value. d Quantitative real-time PCR of transcripts of genes related to amino acid synthesis in intracellular persisters exposed to 50x MIC oxacillin for the indicated periods. Data are means ± SEM of three independent experiments. a, d Source data are provided as a Source Data file.