Fig. 1

ROS alter exosomal miRNA signatures while miR-155-5p being the most downregulated miRNA. A. Exosomes were isolated from A2780 cells by ultracentrifugation. Exosomes (Exo) and total cell lysate (TCL) were analyzed by immunoblotting for CD63, CD81 and Calnexin. B. Exosomes isolated from A2780 cells were analyzed by nanoparticle tracking analysis (NTA). C-E. A2780 cells were treated with or without H₂O₂ at 100 μM for 4 h. 100 ng of exosomal RNAs were used as input for Nanostring nCounter miRNA analysis. Three biological replicates were used. The heatmap of exosome miRNAs profile (C), the volcano plot (D), and the fold changes of representative differentially expressed exosomal miRNAs in treated cells vs. control cells are shown (E). F. The expression levels of cellular and exosomal miR-155-5p were determined in A2780 cells treated with H₂O₂ (100 μM, 4 h). G-H The expression levels of cellular and exosomal miR-155-5p were determined in A2780, SKOV3, and Ovcar-3 cells treated with NAC (10 mM, 4 h). \I. The expression levels of cellular miR-155-5p were determined in A2780 cells treated with catalase-PEG (250 U/ml, 4 h) or rotenone (2.5 μM, 4 h). J. miR-155-5p expression was detected in human ovarian carcinoma tissue microarray (normal N = 40, ovarian cancer N = 160) by in situ hybridization using double DIG-labeled miRCURY LNA miRNA detection probes (red). Data are representative of 3 independent experiments. Data = Mean ± SD. *p < 0.05, **p < 0.01