Fig. 2: Impact of EV inhibitors on EV-mediated increased migration of hSCs.

a hSCs were treated with PKH67-labelled Panc1-derived EVs pre-incubated with different concentrations of heparin or EIPA for 1 h or throughout the experiments (24 h). Images were taken by the Operetta CLS^TM high-content analysis system. Quantification was performed by measuring the intensity of Alexa Fluor 488. The error bars depict mean ± S.E.M. from three independent experiments. b Cell viability of hSCs was measured by PrestoBlue reagent after exposure to Panc1-EVs, heparin or EIPA for 24 h. Data were obtained from three independent experiments and shown as mean ± S.E.M. c 3D migration assay of hSCs mixed with Panc1-EVs pre-incubated with heparin for 1 h before embedding in a Matrigel drop and monitored for 4 days. Images (pseudo-colour) were taken by the digital phase contrast function of the Operetta system. Quantification was performed by measuring the area covered by cells (µm²). Statistical difference of all data was analysed by two-tailed unpaired Student’s t-test. *p < 0.05, **p < 0.01, ***p < 0.001.