Fig. 6: Ag⁺ B cell metabolic states.

A Uniform Manifold Approximation and Projection (UMAP) plot shows the 2D spatial distribution of 9,780 cells from HD and MS patients. UMAP dimensionality reduction is calculated using sampled data from all cells and all available metabolic features; through FlowSOM clustering are identified 8 different clusters, defined as scMEP. Cells are colored by their scMEP state. B Heatmap of the median marker intensities of the 23 metabolic markers across the 8 cell populations, obtained with FlowSOM algorithm. The colors of cluster_id column correspond to the colors used to label the UMAP plot clusters/scMEP. The color in the heatmap is referred to the median of the arcsinh marker expression (0–1 scaled): white represents a lower expression, while dark green represents a higher expression. Light gray bar along the rows (clusters/scMEP) and values in percentages indicate the relative sizes of the clusters. C Projection of UMAP graphs stratified by HD and all different MS patients. D Representative dot plots showing percentages of scMEP1, scMEP3 and scMEP4 among different Group_IDs. The central bar represents the mean ± SD. Kruskal–Wallis test (one-sided) with Benjamini–Hochberg correction for multiple comparisons is used to test the differences among groups. Adjusted q-values are reported in the figure, if significant. healthy donors (HD, n = 8), multiple sclerosis patients treated with Cladribine (n = 4), Dimethyl Fumarate (DMF, n = 8), Fingolimod (n = 5), IFN (n = 6), Natalizumab (n = 8), Teriflunomide (n = 5), Rituximab/Ocrelizumab (n = 7). E Heatmap of 15 immunological markers enrichment modeling (not used for metabolic clustering) across different scMEP states, showing the relationship between metabolic states and functional properties. Light gray bar along the rows (clusters/scMEP) and values in percentages indicate the relative sizes of the clusters/scMEP. F Pseudotime visualization of scMEP development based on the estimated trajectory and envisaged in UMAP space. Colors are representative of the different distribution of cells population: blue represents less active metabolic state, while red represents an increased/enhanced metabolic state.