Fig. 3

MiR-203a-3p of hepatocyte-derived exosomes caused the re-expression of E-cadherin through blocking Src expression. (A) Exosomes derived from LO2 cells were observed by electron microscopy: a. green labelled exosome from LO2 co-cultured with LoVo-P. b. Green labelled exosome co-cultured with LoVo-P under fluorescence microscope. c. Unlabelled exosome from LO2 co-cultured with LoVo-P. d. Unlabelled exosome from LO2 co-cultured with LoVo-P under fluorescence microscope. (B) Wavelengths of exosomes, size distribution of vesicles identified in the 110,000 g medium pellets of 25 vesicles. (C) CD63, HSP70 and TSG101 which are marker proteins in exosomes were examined by western blot. (D) Expression of E-cad and Vimentin in LoVo-P cells co-cultured with LO2 exosoms. (E-F) RNA was extracted from the LO2 cells 110,000 g medium pellet, a human miRNA array was used to screen miRNA expression in LO2-derived exosomes, the front 1000 kurtosis expression of microRNA were used(E), and the filtered microRNA results were confirmed by qPCR (F). (G-H) An exogenous miR-203a-3p mimic (50 nM) was transfected into LoVo-P cells, qPCR (G) and western blot (H) assays were used to examine the expression of Src, p-EGFR and E-cadherin. (I) Predicted binding sites of miR-203a-3p within the 3′-UTR of Src mRNA. (J) Transwell and scratch assays for number of invaded LoVo-P cells co-cultured with LO2 cells exosomes or transfected with the exogenous miR-203a-3p mimic. The data represent the average of three independent experiments. **P < 0.01 compared with control