Inhibition of Egr-1 activation and endothelial cell migration by MβCD.

<p>(A) HMEC-1s were treated with DiI-labeled SW480-derived EVs (1 µg/mL) for 1 h in the presence or absence of MβCD (10 mM) (n = 3). SW480-derived EVs and nuclei were stained with DiI (red) and Hoechst (blue) respectively. Representative photographs are shown in panel A. Scale bars represent 10 µm. (B) Confluent HMEC-1s were scratched and treated with SW480-derived EVs (1 µg/mL) in the presence or absence of MβCD (10 mM); then the number of migrated cells in the denuded zone was evaluated after 12 h (n = 3). (C, D) HMEC-1s were pretreated with MβCD (10 mM) for 1 h and then stimulated with SW480-derived EVs (1 µg/mL) for 1 h. Scale bars represent 30 µm. Nuclear translocation of Egr-1 protein was analyzed using confocal microscopy and the percentage of Egr-1-positive nuclei was determined by measuring the number of cells with nucleus co-localized signals over that of total cells (n = 3). Data are represented as mean ± SD. ***<i>P</i><0.001.