Fig. 3.

Cx43-containing EVs loaded with dox decrease the activation of signalling pathways related to cell motility. The 4T1^luc2 cells were treated with 2 µM free dox, EV^Cx43+ dox, EV^Cx43− dox or vehicles, for 24 h. (a) Time-lapse microscopy was performed to assess cell motility, followed by individual cell tracking. Mean distance and velocity are plotted in the graph (n=3, ^###p<0.001 vs. EV^Cx43+ ; *p<0.05 EV^Cx43+ dox vs. dox;). Left panel shows representative images of the assay. Scale bar, 10 µm. (b) Activation of the p44/42 pathway was analysed by WB. Calnexin was used as loading control (n=3, ^#p<0.05 vs. EV^Cx43+).