Fig. 2: Single-cell metabolic regulome profiles of T cell activation dynamics.
a, Experimental setup to benchmark scMEP mass cytometry analysis with bulk metabolic analysis by extracellular flux analysis (Seahorse). b, Examples of mass cytometry-quantified expression levels of glycolytic (top) and TCA/ETC (bottom) enzymes following no (resting, left) and 3 days (right) of activation of naïve T cells. Examples show data of one representative experiment (out of N = 4 independent experiments). c, Expression levels of important determinants of glycolysis on naïve CD8+ T cells. Black dots indicate population medians. d, Expression levels of TCA/ETC components on naïve CD8+ T cells as in c. e, Extracellular flux analysis of bulk cell populations from the same donor as in a-c. Extracellular acidification rate (ECAR; top) and oxygen consumption rate (OCR; bottom) for each measurement following injections of mitochondrial modifiers. FCCP = fluoro-carbonyl cynade phenylhydrazon, Rot = Rotenone, AA = antimycin A. Shown is data from one individual (out of N = 4 independent experiments). Circles and error bars represent mean±s.d. for three technical replicates (wells). f, Correlation between glycolytic and oxidative enzymes (left panel) and basal glycolysis (top row) or basal respiration (bottom row). Mass cytometry and extracellular flux analysis values were asinh transformed. Circles represent mean population values for each donor and are based on technical replicates (single cells in mass cytometry and three replicate wells in flux analysis). Black lines and r2 values represent results of a linear regression model, with black shading representing the 95% confidence interval (CI). Log10 of (Benjamini-Hochberg; BH) false discovery rate (FDR)-adjusted P-values and r2 values from linear regression models (middle panel). Black line indicates a BH-corrected P-value of 0.05. Protein-based scMEP scores (right) represent the mean expression of all metabolic enzymes within a pathway. Each circle represents the mean scMEP score of a T cell population (naïve or memory). g, Linear correlation of mean (based on three technical replicates) flux analysis values (left) and single-cell scMEP scores (right) of naïve CD8+ T cell populations, calculated as in f. Shown is data from one individual (out of N = 4 independent experiments). Red lines and r2 values represent results of a linear regression model, with black shading representing the 95% CI. h, scMEP scores as in g, visualized for each day.