Fig. 7

The specificity of miR-320a targeting CBL. A A schematic diagram of the cellular model. B si-CBL was transfected into A549 cells. The expressions of CBL was analysed by Western blotting. C A549 cells were co-cultured with EVs (WT-EVs, miR-320a KO-EVs), and the expression of miR-320a was detected by qRT-PCR. D qRT-PCR was used to detect the mRNA levels of CBL after treatment with EVs (WT-EVs, miR-320a KO-EVs). E A549 cells were treated with EVs (WT-EVs, miR-320a KO-EVs), and then cells were collected for Western blotting to show CBL expression. F Changes in cellular oxidative phosphorylation were detected by extracellular flux analysis. G and H ATP production and mtDNA expression in A549 cells after treatment with EVs (WT-EVs, miR-320a KO-EVs). I Mito Tracker Red staining of mitochondria in A549 cells. Scale bar, 20 μm. Mito Tracker Red: red, DAPI: blue. J The quantitative analysis of mitochondrial fluorescence intensity. K The mitochondrial potential was observed via JC-1 staining. L The red to green fluorescence ratio was recorded to quantify the mitochondrial potential (rate). Scale bar, 20 μm. M ROS levels were assessed. N The relative fluorescence intensity ratio shown in (M) was analyzed. Scale bar, 20 μm. O qRT-PCR was performed to determine mRNA levels of Drp1, Mfn1, and Mfn2. P CCK-8 assay was used to determine the Cell proliferation. Q Caspase 3 activity was measured in the control, miR-320a mimics, and miR-320a inhibitor groups. R The expressions of Caspase 3, Bcl-2, and Bax were analyzed by Western blotting. 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