Figure 4. Transcriptomic visualization of thermogenic adipocytes in iWAT.

(A) Aggregated tSNE-plot showing 17 clusters from ~54,000 adipocytes derived from iWAT of mice at RT, cold-exposed (24 hr, 48 hr, and 4 days), or treated with CL for 4 days at 1 mg/kg/day. Each colored dot is an adipocyte that falls into a cluster based on transcriptomic signature. (B) tSNE-plot showing indicated treatment-dependent sub-clustering of aggregated clusters shown in (A). Black arrow indicates the Type nine adipocyte cluster. (C) Normalized expression values of the top two adipocyte subtype-specific cluster genes from (A and B) plotted as violin plots with clusters as rows and genes as columns. (D) tSNE-plot showing distribution of indicated genes from adipocytes from (A). (E) Normalized expression values of indicated genes in the Type nine adipocyte cluster under different treatment condition plotted as violin plots with treatment conditions as rows and genes as columns. (F) RNAScope FISH (see Materials and methods) of indicated probes from iWAT of mice treated with saline or CL for 4 days at 1 mg/kg/day.

Figure 4—figure supplement 1. Enrichment of cells involved in lipid mobilization in Cluster nine adipocytes.

(A and B) Gene set pathways obtained from GO Biological Processes and KEGG were used to find the functions of the different cell types. The top five enriched pathways determined by FDR from each cell type cluster were selected (from Supplementary file 1) and only unique pathways were kept (some top enriched pathways were shared across cell type clusters). Enriched pathways (rows) were clustered with hierarchical clustering. The size of each dot represents the -log10 FDR of the pathway enrichment and the color of each dot corresponds to the fold enrichment of each pathway (red- higher enrichment, gray- lower enrichment). Dotted blue squares highlight pathways enriched in cluster 9.

Figure 4—figure supplement 2. Type nine cluster constitute of thermogenic adipocytes.

(A) Normalized expression values of top adipocyte subtype-specific major clusters (1, 4, and 9) genes from Figure 4A plotted as violin plots with clusters as rows and expression levels of genes as columns. (B) tSNE-plot showing cluster-specific expression of selected marker genes from aggregated tSNE-plot in Figure 4A. Black arrow indicates Type nine cluster. (C) Normalized expression values of indicated genes plotted as violin plots with clusters as rows and expression levels of genes as columns.