Fig. 2: EECs-derived exosomes deliver miR-30c to repress the invasion and migration of ecto-EECs.

A TEM observation of exosomes derived from ecto-EECs and normal EECs; B Nanoparticle tracking analysis of the size and distribution of exosomes derived from ecto-EECs and normal EECs; C Expression levels of CD9, CD63, and HSP70 in isolated exosomes and cell extracts, determined by Western blot assay; D The expression of miR-30c in exosomes derived from ecto-EECs and normal EECs, determined by qRT-PCR; E Immunofluorescence detection of ecto-EECs cocultured with PKH26-stained exosomes derived from normal EECs (Red indicates exosomes and blue indicates DAPI-stained nuclei); F The expression of miR-30c in exosomes derived from miR-30c-inhibiting normal EECs, determined by qRT-PCR; G The expression level of miR-30c in ecto-EECs following coculture with exosomes from miR-30c-inhibiting normal EECs, determined by qRT-PCR; H Wound healing assay to detect cell migration in ecto-EECs following coculture with exosomes from miR-30c-inhibiting EECs, reflected by quantification of the wound healing rate; I Transwell assay to detect cell invasion in ecto-EECs after coculture with exosomes from miR-30c-inhibiting EECs, with the number of invasion cells counted; J Nanoparticle tracking analysis of the size distribution and number of exosomes derived from GW4869-treated normal EECs (GW4869, an exosome release inhibitor); K Wound healing assay to detect cell migration in ecto-EECs following coculture with exosomes from GW4869-treated normal EECs, reflected by quantification of the wound healing rate; L Transwell assay to detect cell invasion in ecto-EECs after coculture with exosomes from GW4869-treated normal EECs, with the number of invasion cells counted; M Expression levels of EMT-related proteins in ecto-EECs cocultured with exosomes from EECs of different groups, determined by Western blot. Cellular experiments were repeated three times. Measurement data were characterized as mean ± SD. The comparison between data of two groups was performed by independent sample t-test, and that among multiple groups was performed with one-way ANOVA with Tukey’s post hoc test. *p < 0.05 versus the Normal group, inhibitor-NC group, PBS group, or DMSO group; #p < 0.05 versus the Normal exosome group; &p < 0.05 versus the (Normal + DMSO) exosome group.