Extracellular vesicles(EVs)were isolated from the culture supernatant of lung adenocarcinoma A549 cells.The size of EVs was characterized as approximately 100 nm via DLS and TEM analysis,with the EV-specific marker TSG101 detected in the samples.The extracted EV DNA(evDNA)showed a predominantly～12 kb size distribution as assessed by agarose gel electro-phoresis.Based on sequencing data,specific evDNA was selected for PCR detection.DNase I or PS nuclease treatment of the EVs confirmed that the evDNA existed on the outside of the vesicle in a linear manner,and poly-dA tails were added to evDNA by TdT enzyme treatment,allowing for their capture by oligo-dT beads and presenting further evidence of their linear surface distribution.Additionally,a TetO-DNA/TetR-GFP visualization system was established to confirm the presence of TetO-DNA in the EVs,which could be visualized by TetR-GFP proteins under fluorescent microscopy.Enrichment of EVs carrying the TetO-DNA was confirmed via flow cytometry and PCR experiments using TetR-GFP protein.These findings collectively suggest that evDNA is distributed linearly on the surface of EVs,providing a basis for further research into their biological functions.
国家科技期刊平台
NSTL
维普
万方数据
