Fig. 7

SK1-KO EV failed to activate HSC and promote FMC metastasis to liver. A SK1-KO EV failed to activate HSC. LX-2 cells were incubated with PBS (control), FMC-EV, or SK1-KO EV for 24 h. The expression of α-SMA in LX-2 cells was analyzed by Western blotting. The samples were analyzed in triplicate. Protein band intensity was quantified using ImageJ (right panel) B SK1-KO EV failed to activate STAT3 in HSC. Flow cytometric analysis of the p-STAT3 level in LX-2 cells after treatment with PBS (control), FMC-EV, or SK1-KO EV. The mean fluorescence intensity of p-STAT3 is shown (right panel). n = 3 in each group. C SK1-KO EV failed to promote migration of HSC. LX-2 cells incubated with PBS (control), FMC-EV, or SK1-KO EV were grown in the Culture-Insert (ibidi) for a wound healing assay. Images were acquired after 48 h, and the closure area of the wound was quantified using ImageJ. n = 3 in each group. D Immunohistochemical analysis of α-SMA (red) in livers taken from NPG mice intraperitoneally injected with PBS (control), FMC-EV, or SK1-KO EV for three weeks. The liver sections were counterstained with hematoxylin (blue). E Representative results of ex vivo imaging of the livers harvested from the mice intraperitoneally injected with PBS (control), FMC-EV, or SK1-KO EV, followed by intrasplenic injection with FMC-1807-RFP. The fluorescence intensity of FMC-1807-RFP in liver was acquired as radiant efficiency. n = 6 in each group. Quantitative data are presented as the mean ± SD. The Student’s t-test was used to determine the statistical significance