Extended Data Fig. 3: GLS depletion promotes metabolic gene programs independently of effects on adipogenesis.
From: Reduced adipocyte glutaminase activity promotes energy expenditure and metabolic health
a-c. In vitro differentiated human adipocytes were transfected with siC or siGLS. Effects on (a) gene expression (four replicates per condition, repeated >three times), (b) protein abundance (repeated twice) of adipogenic markers (left) and adiponectin secretion (in total sixteen replicates per condition from two independent experiments) (right) and (c) lipid droplet morphology (repeated >three times) were determined. Scale bar in panel c = 20 μm. Proteins from the same samples were loaded on four different gels in parallel. d. Gene set enrichment analysis listing the top five pathways altered in siGLS vs. siC human adipocytes. e. Heatmaps of indicated pathways and genes altered in siGLS vs. siC treated human adipocytes. f. Representative examples of electron transport chain (ETC) protein levels in human adipocytes incubated with 0, 2.5 and 10 mmol/L of glutamine. Cells were either deprived or treated with indicated concentrations of glutamine for 48 hours (repeated three times). Proteins from the same samples were loaded on two different gels in parallel. g. Representative examples of ETC protein levels in human adipocytes transfected with siC or siGLS oligonucleotides incubated with or without eta-ketoglutarate (EtaKG, repeated three times). h. Representative examples of ETC protein levels in human adipocytes transfected with siC or siGLS oligonucleotides and with scrambled mRNA or over expressing GLS mRNA. (repeated two times). Error bars in panels a-b are S.E.M. Abbreviations: ADIPOQ = adiponectin, BODIPY = boron-dipyrromethene, CEBPA = CCAAT/enhancer-binding protein alpha, PLIN1 = Perilipin 1, PPARG = peroxisome proliferator-activated receptor gamma.
