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. 2019 Aug 19;15(10):2240–2255. doi: 10.7150/ijbs.35356

Figure 4.

Figure 4

miR-27a-containing exosomes secreted by irradiated HaCaT cells mediate bystander effects in unirradiated WS1 cells. (A) The representative TEM images of the exosomes isolated from media culturing unirradiated and irradiated HaCaT cells. (B) The concentrations of miR-27a/b in the exosomes collected from unirradiated and irradiated HaCaT cells at different times post radiation. (C) Uptake of exosomes from unirradiated and irradiated HaCaT cells by the recipient WS1 cells. Exosomes were pre-labelled with DiI, cell nuclei were stained with DAPI. (D) The alterations of the intracellular ROS levels of the recipient WS1 cells after culture with the exosomes from unirradiated and irradiated HaCaT cells collected at different times post radiation for 1 h, showing that the exosomes from irradiated HaCaT cells collected at 3 and 6 h but not 12 h post radiation caused oxidative stress in the recipient WS1 cells. (E) The quantification of area of the wound scratches of the recipient WS1 cells after culture with the exosomes from unirradiated and irradiated HaCaT cells collected at different times post radiation, suggesting that the exosomes from irradiated HaCaT cells collected at 3 and 6 h but not 12 h post radiation slowed the migration of the recipient WS1 cells. (F) The concentrations of miR-27a in the 3 h exosomes from unirradiated and irradiated HaCaT cells pre-transfected with miR-27a inhibitors and NC, showing lack of increase in miR-27a in the exosomes from irradiated HaCaT cells with down-regulated miR-27a. (G) The alterations of the intracellular ROS levels of the recipient WS1 cells after culture with the 3 h exosomes from unirradiated and irradiated HaCaT cells pre-transfected with miR-27a inhibitors and NC, showing that down-regulated miR-27a in exosomes significantly inhibited the oxidative stress in the recipient WS1 cells. (H) The quantification of the area of the wound scratches of the recipient WS1 cells after culture with the 3 h exosomes from unirradiated and irradiated HaCaT cells pre-transfected with miR-27a inhibitors and NC, showing that down-regulated miR-27a in exosomes significantly inhibited the slowed migration of the recipient WS1 cells. All the data represent the means ± SEM from three independent experiments (n=3). *P<0.05, and **P<0.01 compared with the relative control. NS, not significant.