Fig. 7: Lipolysis activates overlapping gene programs in human brown adipocytes.

a Heatmap of RNA-seq data showing lipolysis-activated genes (Log2FC ≥ 0.7, FDR/Benjamini–Hochberg ≤ 0.05) in human in vitro differentiated brown adipocytes. Cells were stimulated with 20 µM SR-3420 or vehicle for 3 h before harvest. n = 3 biologically independent experiments examined, each carried out in a technical duplicate. b Pathways significantly enriched (FDR ≤ 0.05) among lipolysis-activated genes in a. c Examples of lipolysis-activated genes derived from RNA-seq belonging to enriched pathways in b. d Heatmap of RNA-seq data showing dependency of lipolysis for induction of genes by ISO Log2FC ≥ 1, FDR/Benjamini-Hochberg ≤0.05) in white adipocytes. Mature mouse white adipocytes were isolated from epididymal white adipose tissue and cultured in a 3D matrix for 2 days. After 2 days, cells were pre-treated with 10 µM Atglistatin (ATGL inhibitor) and 20 µM CAY10499 (HSL inhibitor) for 1 h and subsequently with 20 µM SR-3420 for 3 h before harvest. n = 3 biologically independent experiments examined, each carried out in a technical duplicate. e Pathways significantly enriched (FDR ≤ 0.05) among lipolysis-dependent and -independent ISO-activated genes in d. f Examples of lipolysis-dependent and -independent ISO-activated genes derived from RNA-seq in d. n = 3 biologically independent experiments examined, each carried out in a technical duplicate. g Venn diagram illustrating the overlap of lipolysis-dependent genes in mouse white (d) and brown adipocytes (Fig. 1d). For all panels, error bars represent ±SEM of 3 independent biological experiments. Statistical significance was determined by using FDR/Benjamini-Hochberg correction for Fig. 1f (p ≤ 0.05 = *, p ≤ 0.01 = **, p ≤ 0.001 = ***). * versus Vehicle, # versus ISO (without Lipase inh.).