Fig. 2: The correlation analysis of intracellular metabolome and cellular OS level.

a The diagram illustrates the pairing of metabolic data and OS levels, as well as the subsequent correlation analysis. The DCFDA intensity indicated the specific level of OS in each cell, while single-cell MS was employed to determine the abundance of metabolites. Here, the metabolite GSH in Cells 1–4 was used as an illustrative example. Data from Cell 1–4 was normalized to Cell 1. The vectors for x and y were constructed based on single-cell OS levels and metabolite abundances, followed by calculation of Pearson’s coefficient (r) and significance (P). In correlation analysis, the data was z scored. Scale bar, 15 μm. b Metabolites that were inversely and positively correlated with the level of OS. Metabolites are shown as dots by color: red, positively correlated (P < 0.05); blue, inversely correlated (P < 0.05). The representative metabolites are annotated adjacent to the corresponding dots. P values were not adjusted. Two-sided Pearson’s correlation analysis was performed. c–h The correlation analysis between the scaled levels of OS and the scaled intensities of representative metabolites including ATP (c), PCr (d), UTP (e), GTP (f), Hypt (g), and energy charge (h). The data were standardized by calculating z score. P values were not adjusted. Two-sided Pearson’s correlation analysis was performed. i The metabolic pathways enriched by the MSEA analysis of metabolites exhibiting a reverse correlation with OS levels. Only metabolites with MS/MS confirmation were included in the analysis. Only pathways with P < 0.05 in the MSEA analysis were included. P values were not adjusted. Related metabolic processes are annotated on the left. Dot size represents enrichment ratio, and dot color represents significance of the enrichment (P value). Yellow, relatively high significance; Blue, relatively low significance. n = 190 cells in (a) and (c–h). For (a–h), Source data are provided as Source Data files. ATP adenosine triphosphate, PCr phosphocreatine, UTP uridine triphosphate, GTP guanosine triphosphate, Hypt hypotaurine, GSH glutathione, CTP cytosine triphosphate, O-PE O-phosphoethanolamine, G-3-P glycerol 3-phosphate, AMP adenosine monophosphate.