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. 2021 Aug 27;22(17):9289. doi: 10.3390/ijms22179289

Figure 6.

Figure 6

(20S)G-Rh2 inhibited Anxa2-Y23D mutant-enhanced proliferation and migration of HUVECs. Myc-tagged wild-type, Y23A mutant and Y23D mutant of Anxa2 was over-expressed in HepG2 cells. (A) The concentration of four VEGFs in cell-free supernatant of HepG2 cells was determined by ELISA. (B,C) Colony formation of HUVECs was performed with cell-free supernatant from HepG2 cells pre-treated with (20S)G-Rh2 or not. The cell amount of each colony was shown as superimposed symbols with connecting lines (B), and the overall status of colony formation was shown as images after crystal violet staining(C). (D,E) EdU-labeling was utilized for new DNA synthesis detection in HUVECs co-cultured with HepG2 cells. EdU-positive cells were shown as florescent images (D), and EdU-positive ratio was shown as histogram (E). (F) Migrated HUVECs co-cultured with HepG2 cells were shown as images after crystal violet staining. All experiments were performed with triple replicant. Significance analysis was shown *** (p < 0.001). Scale bar presented 20 μm.