Fig. 5.

Studying the presence of lipids in adipose tissue (AT)-extracellular vesicles (EVs) by confocal microscopy (a). Confocal microscopy analysis was performed for Simpson Golabi Behmel Syndrome (SGBS) adipocyte and patient AT-derived EVs that were stained for lipids and CD63. EV samples from pre- and mature SGBS cells, as well as from patient visceral and subcutaneous adipose tissue (VAT and SAT, respectively) ex vivo cultures were stained with LipidSpot lipid droplet stain and fluorophore-conjugated CD63-antibody, after which samples were imaged with high-resolution confocal microscopy. PBS was included as non-EV control. The mRNA expression levels of PNPLA2 in SGBS cells treated with 20 ng/ml TNFα for 24 h were analyzed by RT-qPCR (b). The values of six independent experiments are shown, presented as mean + SEM. **p = 0.002 (Mann–Whitney U test). Phosphorylated levels of hormone sensitive lipase (HSL) were studied by Western Blotting, from three independent experiments (c). Results are presented as mean + SEM. The mRNA expression levels of LIPA were studied by RT-qPCR, from nine independent experiments (d). Results are presented as mean + SEM, ***p = 0.0001 (Mann–Whitney U test). Glycerol concentration was determined from culture media of three experiments (e), and Rab7 protein levels from cells of four experiments (f). Results are presented as mean + SEM. *p = 0.014 (Mann–Whitney U test)