Figure 7
From: Antibiotic-induced release of small extracellular vesicles (exosomes) with surface-associated DNA
Assessment of DNA and DNA-binding proteins in extracellular vesicle (EV) preparations. (a-b) DNA was purified from apoptotic bodies (APOs), microvesicles (MVs) or exosomes (EXOs) released by ciprofloxacin-exposed control, activated or apoptotic Jurkat cells. (a) Nuclear (GAPDH, p53) and (b) mitochondrial (control region, RNR1) DNA sequences of DNase I-digested/non-digested EVs were amplified by PCR and analyzed by agarose gel electrophoreses. The figure displays cropped gels. Full-length, uncropped gels are shown in Supplementary Figures S11–S13. (c ) Detection of exosomal DNA derived from ciprofloxacin-exposed control, activated or apoptotic Jurkat cells using an Agilent 2100 Bioanalyzer (DNA 12,000 Kit). The electropherograms show the size distribution of purified exosomal DNA in base pairs (bp) with DNA markers at 50 bp and 17,000 bp. FU: fluorescence units. (d) Semi-quantitative mass spectrometry analysis of DNA-binding histones in EV samples. Values in the table are proportional to the amount of histones found in EVs. The presence of f lap endonuclease 1 (FEN1, also known as a mitochondrial DNA-binding nucleoid protein) was also identified by mass spectrometry.
