Fig. 1

Aging of human adipose tissues is accompanied by the appearance of PLAU⁺ adipose progenitor cells and dominating role of macrophages. a t-Distributed stochastic neighbor (t-SNE) plot shows unsupervised clustering of 13841 single-cell transcriptomes of SVF cells from 3 young individuals and 3 old individuals. b Violin plots show the linage marker genes expression level in all cell clusters. c Scatter plot shows the changes in the proportion of 7 APC subpopulations during the aging process. Data are presented as the mean ± s.d. of three young and three old individuals. d Dot plot shows the expression levels of chemokine activity-related genes and adipogenic-related genes across 7 APC subpopulations of 3 young and 3 old individuals. e t-SNE plots show the expression level of THBD and PLAU in all APC clusters in young and old individuals. f Representative images of immunofluorescence staining of PLAU and THBD in human SAT of young and old group. Scale bar, 50 μm. g FACS analysis of PLAU⁺, THBD⁺ APC in SAT of young and old group. h UMAP visualization of all APC subpopulations. RNA-velocity analysis with velocity field projected onto the UMAP plot of APC subpopulations, arrows show the local average velocity evaluated on a regular grid and indicate the extrapolated future states of cells (top). Cells were annotated by monocle pseudotemporal dynamics (purple to yellow, below). i, j APC populations of the second cohort. i Scatter plot shows the changes in the proportion of 6 APC subpopulations during SAT aging process. Data are presented as the mean ± s.d. of 4 young and 4 old individuals from the second cohort. j Dot plot shows the expression levels of chemokine activity-related genes and adipogenic-related gene across 6 APC subpopulations of 4 young and 4 old individuals from the second cohort. k qRT-PCR for PLAU expression level of human APC after PLAU knockdown. Data are presented as the mean ± s.d. of three biological repetitions. l Representative images of SA-β-gal staining of human APC upon shRNA-mediated knockdown of PLAU. Scale bar, 50 μm. Positive signals were quantized by average optical density (AOD). Data are presented as the mean ± s.d. of 3 fields of each group. m Representative images of Oil Red staining shows the adipogenic differentiation capacity of human APC upon shRNA-mediated knockdown of PLAU. Scale bar, 50 μm. Data are presented as the mean ± s.d. of 3 cell wells of each group. n Representative images of immunofluorescence staining of Ki67 in human APC upon shRNA-mediated knockdown of PLAU. Scale bar, 50 μm. Data are presented as the mean ± s.d. of 4 fields of each group. o Re-clustering of IC1-IC3 in a, b identified 8 specific immune cell subpopulations, samples from 3 young individuals and 3 old individuals. p Dot plot shows the expression levels of representative cell-type-specific marker genes across all these 8 immune cell subpopulations. q Interaction heatmap plots the total number of cell receptor (y axis) and ligand (x axis) interactions in 3 young donors (left) and 3 old donors (right) derived SAT. The color key from blue to red indicates low to high number of interactions. r Circos plots shows the top 20 chemokines mediated ligand-receptor interaction for all APC and immune cell subpopulations (left, young group, n = 3; right, old group, n = 3). s–u Primary human APC treated with conditioned media from DOX-induced senescent macrophages. s Representative images of SA-β-gal staining of conditioned media treated APC. Scale bar, 50 μm. Data are presented as the mean ± s.d. of 3 fields. t Representative images of Oil Red staining show the adipocyte differentiation of conditioned media treated APC. Scale bar, 50 μm. Data are presented as the mean ± s.d. of 3 biological replications. u Representative images of immunofluorescence staining of Ki67 in conditioned media treated APC. Scale bar, 50 μm. Data are presented as the mean ± s.d. of 3 fields. YSAT, young subcutaneous adipose tissues; OSAT, old subcutaneous adipose tissues; -DOX CM, conditioned medium of macrophages without DOX treatment; +DOX CM, conditioned medium of macrophages treated by DOX. In dot plot (d, j, p), the size of the dot corresponds to the percentage of cells expressing the specific gene in each cluster, and the color encodes the scaled average expression level of feature genes across all cells within a subpopulation. *P < 0.05; **P < 0.01; ***P < 0.001; ***P < 0.001