Fig. 2.

Studying extracellular vesicle (EV) secretion and EV fatty acid (FA) profiles from adipocyte treatments. Mature Simpson Golabi Behmel Syndrome cells were treated with either 20 ng/ml of tumor necrosis factor α (TNFα), 400 µM of palmitic acid (PA, 16:0) or 75 µM eicosapentaenoic acid (EPA, 20:5n-3) for 24 h, after which EVs were isolated and analyzed by nanoparticle tracking analysis (NTA). Both particle counts (a) and size distribution of particles (b) were obtained by nanoparticle tracking analysis NTA. Particle counts have been normalized to cell number, and results are presented as mean + SEM. *p < 0.05 (Mann–Whitney U test). Differences in FA profiles of cells and secreted EVs from TNFα, PA, and EPA treatments were determined from total lipids with gas chromatography–mass spectrometry (c). Results are presented as percentage differences, calculated by subtracting the mol-% of each FA in the control group from the mol-% in the treatment group. Red indicates an increase in the treatment group, while blue indicates a decrease. FAs are listed in the order of increasing chromatographic retention time. DMA plasmalogen alkenyl chain-derived dimethyl acetal derivative, SFA saturated fatty acid, MUFA monounsaturated fatty acid, PUFA polyunsaturated fatty acid, unsaturated FA (UFA) = MUFA + PUFA. *p ≤ 0.05 Mann–Whitney U test vs. control. Percentages of selected FAs in cells and secreted EVs from TNFα, PA, and EPA treatments, presented as mean mol-% (d). The FA results of TNFα and PA treatments were measured from 5 independent experiments and the results of EPA treatments from 7 independent experiments. The supervised discriminant analysis depicts the classification of FA signatures of cells and EVs from TNFα, PA, and EPA experiments based on discriminant functions 1 and 2 (e). Function 1 (on the x-axis) explained 81.7% of the variance in the dataset, and Function 2 10.6% of the variance