Extended Data Fig. 1: Related to Fig. 1.
A) Gating strategy for neutrophils and monocytes/macrophages of unfractionated bone marrow from aspirates for single-cell RNA sequencing experiments B) Gating strategy for quantification of mature neutrophils, monocytes and macrophages in unfractionated bone marrow aspirates. Fractions were calculated as percentages of ‘live cells’-gate (as depicted in (A)) C) UMAP of the combined dataset of 47,497 cells from NDMM patients and 34,531 cells from controls showing 16 clusters identified by marker genes in (D) and (E). Colors represent clusters D) Marker gene transcription to demarcate the 3 major myeloid populations: progenitors (MPO+), mononuclear phagocytes (CD14+) and neutrophils (MME+ marks mature neutrophils) E) Transcription of marker genes of the myeloid dataset F) Monocle 3 pseudotime analysis of the myeloid dataset. ‘S’ represents an arbitrarily selected starting point of trajectory calculations G) Cluster distribution of myeloid clusters per condition. Colors represent clusters H) Cluster distribution of myeloid clusters per patient. Colors represent clusters. ‘MM’, MM-patient. ‘CBM’, control I) Chord diagram visualizing frequency of interactions between MSC subsets from 4 and myeloid subsets of MM patients J) Predicted ligand–receptor interactions between MSC clusters from 4 and myeloid clusters from MM patients. Expression magnitudes and interaction specificity values were calculated using the statistical framework of liana22 K) Cluster distribution of neutrophil clusters per patient. Colors represent clusters. ‘MM’, MM-patient. ‘CBM’, control L) Percentages of neutrophils in the 6 remaining clusters per individual in each condition M) Transcription of interferon-response genes comparing non-cancer controls to NDMM patients FSC, forward scatter. SSC, sideward scatter. GMP, granulocyte-monocyte progenitor. Data are presented as mean ± SEM. Significance was calculated in L using the Mann–Whitney U test (two-tailed). NS P > 0.05.
