Fig. 3

EVs released by ADSCs alleviate mitochondrial dysfunction and apoptosis. A EVs from cells (ADSCs, 293 T cells) were extracted and identified by TEM. Scale bar, 100 nm. B EVs detection by NTA in FBS-free media. C Positively expressed EV markers CD63 and CD9 were detected by Western blotting. D Changes in cellular oxidative phosphorylation were detected by extracellular flux analysis. E The generation of ATP in different groups. F Assay of mtDNA expression in A549 cells. G Mito Tracker Red staining of mitochondria in different groups. Scale bar, 20 μm. Mito Tracker Red: red, DAPI: blue. H The quantitative analysis of mitochondrial fluorescence intensity. I The mitochondrial potential was observed via JC-1 staining. J The red to green fluorescence ratio was recorded to quantify the mitochondrial potential (rate). Scale bar, 20 μm. K ROS levels were assessed. L The relative fluorescence intensity ratio shown in (K) was analyzed. Scale bar, 20 μm. M qRT-PCR was performed to determine mRNA levels of mitochondrial fragmentation-related genes, including Drp1, Mfn1, and Mfn2. N Cell proliferation was determined by the CCK-8 assay. O Caspase 3 activity was measured in the control and EVs treated groups. P The expression levels of Caspase 3, Bcl-2, and Bax were analyzed by Western blotting. TEM, fluorescence images and blots were representative of six independent experiments. All data are presented as the mean ± SEM of n = 6. ***p < 0.001, **p < 0.01, *p < 0.05, ns, no significance