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. 2022 Sep 2;13:5184. doi: 10.1038/s41467-022-32849-1

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — A Graphical overview of the scMEP approach depicting metabolic enzymes, signaling factors and metabolite receptors spanning multiple metabolic pathways as well DC-lineage markers profiled by CyTOF. B tSNE visualization of DC stages using scMEP metabolic markers. Heatmap overlay of single-cell CD14, HLA-DR, CD86 and CD1c (arcsinh transformed) expression highlights associations between DC stages and immune marker expression. Data show one representative experiment (out of N = 3 donors). C Shown are (arcsinh transformed) expression values for selected scMEP immune markers across DC differentiation states. Black dots represent population medians and the dotted line separates early precursors from iDC, actDC, and mDC stages. Violin plots are representative of 1 donor (out of N = 3). On the right side are represented summary (mean) kinetic expression profiles for all measured immune and metabolic scMEP parameters across DC differentiation. D Kinetic profiles of protein synthesis-adjusted SCENITH parameters (calculated as described in materials and methods) to obtain metabolic pathway-dependent changes accounting for ATP production. Lines highlight mean SCENITH profiles (precursor stages represent 3 independent donors, iDC, actDC, and mDC represent six independent donors). E Kinetic profiles for calculated mean scMEP pathways scores are illustrated. Connecting lines visualize mean pathway changes (N = 3 donors). F Correlations between median SCENITH parameters and respective calculated median scMEP pathway scores with Spearman correlation coefficient (R), p-value and grey shading denoting 95% confidence interval (CI). Middle and multi-panel graphs depict single-cell scMEP scores for combined and individual DC sample time points respectively. Subsampled single-cell data points for the individual donor (out of N = 3) are shown. Source data are provided as a Source Data file.