Fig. 3: Lipolysis-activated genes are induced by ISO by PPAR-dependent as well as -independent mechanisms.

Mouse in vitro differentiated brown adipocytes were treated with siRNA against Pparg, Ppara, or NT (negative control) for 3 days and subsequently with 100 nM isoproterenol (ISO) for 3 h before harvest. a Model indicating the potential role of PPARs in the transcriptional activation of genes in C1, C3 and C4 by β-adrenergic signals. Created with BioRender.com. b mRNA expression of adipocyte genes with or without Pparg knockdown in mature brown adipocytes quantified using qPCR. n = 2 biologically independent experiments examined, each carried out in a technical duplicate. c Micrograph of mature brown adipocytes with or without Pparg knockdown. Pictures are representative of n = 2 biologically independent experiments examined. Scale bar denotes 10 µM. d mRNA expression pattern of representative PPAR-dependent and -independent genes derived from RNA-seq after stimulation with 100 nM ISO for 3 h. n = 2 biologically independent experiments examined, each carried out in technical duplicates. e Pathways significantly enriched (FDR < 0.05) among PPAR-dependent and -independent genes in mouse-brown adipocytes stimulated with 100 nM ISO for 3 h. For all panels, error bars represent ±SEM of 2 independent biological experiments. Statistical significance was determined by DESeq2 using FDR/Benjamini-Hochberg correction for b, c (p ≤ 0.05 = *, p ≤ 0.01 = **, p ≤ 0.001 = ***). * versus siNT, # versus Vehicle.