Fig. 6
SERPINE1 was enriched in EVs secreted from STCs. (A) Venn diagram illustrating the similarities and differences between our study and ExoCarta and Vesiclepedia databases. (B) Venn diagram showing overlaps between our study and the SASPAtlas. (C) Venn diagram depicting the proteins detected in our study and those detected from the HCT116 cell line in the NCI-60. (D) Volcano plot of differentially expressed proteins between Ctrl-EVs and Sen-EVs. (E and F) CRC cells were treated with CPT-11 for 96 h to induce senescence, and untreated CRC cells were used as a control. (E) SERPINE1 protein level in whole cell lysates and EVs. (F) SERPINE1 mRNA level in CRC cells. (G) EVs derived from STCs were treated with proteinase K in the presence or absence of Triton X-100 and analyzed by western blot. (H) Scheme of cells cultured with Sen-EVsSERPINE1−GFP. CRC cells were plated in dishes and transfected with GFP-SERPINE1. After 24 h, cells were treated with CPT-11 for 96 h to induce senescence. The cultured cells were washed twice with PBS and cultured in serum-free DMEM for 48 h, then the culture media was collected for Sen-EVsSERPINE1−GFP isolation. The recipient cells were co-incubated with the Sen-EVsSERPINE1−GFP (15 µg/ml) for 8 h and photographed under a fluorescent microscope. (I) Fluorescence imaging of CRC cells treated with Sen-EVsSERPINE1−GFP. (J) SERPINE1 protein level in Sen-EVs and Sen-EVsSERPINE1−GFP. (K) CRC cells were incubated with EVs (15 µg/ml) or PBS for 48 h and the protein expression of SERPINE1 was detected. Data represented the mean ± standard deviation of at least 3 independent experiments. *p < 0.05, **p < 0.01, and ***p < 0.001