Fig. 5: S. aureus simultaneously utilizes lactate and glucose in blood.

a The read counts for all TA sites within the ccpA gene detected by sequencing in both forward (red) and reverse (blue) directions were plotted using R. For identification of the role of the ccpA gene for bacterial growth in blood, ΔccpA strains generated in (b) HG003 and (c) USA300 strains were cultured in blood for 24 h and were counted for CFU. WT; wild-type. For the measurement of lactate and glucose consumption of USA300 in blood and TSB, bacteria were cultured in (d) blood or (e) TSB for up to 36 h. At designated time points, blood and TSB samples were centrifuged for the collection of plasma and supernatant, respectively, which were prepared for mass spectrometric analysis. f USA300 was cultured in human whole blood for up to 36 h for the measurement of amino acid consumption. At designated time points, blood was centrifuged for plasma collection and prepared for mass spectrometric analysis (others include Ala, Arg, Asn, Glu, Gln, His, Leu, Lys, Met, Phe, Pro, Thr, Trp, Tyr, and Val). g For the measurement of bacterial bacillithiol, USA300 was inoculated into human whole blood or TSB for 9, 12, or 24 h. At designated time points, samples were centrifuged for the collection of bacterial cells, after which the bacterial cells were quenched, lysed by bead beating, and prepared for mass spectrometric analysis. Bacillithiol levels at each time point were normalized to log₁₀ bacterial CFU. Glucose, lactate, amino acid concentrations, and bacillithiol levels in d–g were measured using LC/MS-TOF, and the abundance of extracted metabolite ion intensities was acquired using Profinder 6⁶⁰. All data in d–g are presented as the mean ± SD (n = 3 biological replicates). Source data is available in the Source Data file.