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. 2020 Jun 28;11(17):5106–5117. doi: 10.7150/jca.45822

Figure 5.

Figure 5

MST4 regulates HCC cell proliferation and cell cycle by affecting PI3K/AKT pathway. (A) Western blot analysis of AKT, p-AKT, GSK3β and p-GSK3β in MST4-overexpressing Bel-7402 and Bel-7404 cells. (B) Western blot analysis of AKT, p-AKT, GSK3β and p-GSK3β in dnMST4-expressing Bel-7402 and Bel-7404 cells. (C) Western blot analysis of AKT, p-AKT, GSK3β and p-GSK3β in dnMST4-expressing Bel-7402 cells and dnMST4-expressing Bel-7402 cells treated with LY294002 for 24 hours. (D) Cell proliferation of dnMST4-expressing Bel-7402 cells and dnMST4-expressing Bel-7402 cells treated with LY294002 detected by CCK-8 assay (**P < 0.01, Student's t test). (E-F) The anchorage-independent growth of dnMST4-expressing Bel-7402 cells and dnMST4-expressing Bel-7402 cells treated with LY294002 under normoxia and hypoxia was interrogated by soft agar assay, and the statistical results were shown in pane (F) (**P < 0.01). (G-H) The percentage of proliferating cells in dnMST4-expressing Bel-7402 cells and dnMST4-expressing Bel-7402 cells treated with LY294002 was determined by EdU incorporation assay, and the statistical results were shown in pane (H) (*P < 0.05). Scale bar, 500μm. (I-J) Cell cycle distributions in dnMST4-expressing Bel-7402 cells and dnMST4-expressing Bel-7402 cells treated with LY294002 under normoxia and hypoxia were detected by PI staining in triplicate followed by flow cytometry, and the statistical results were shown in pane (J) (**P < 0.01). (K) A schematic diagram of the signal pathway that MST4 inhibits HCC cell proliferation and induces cell cycle arrest via suppression of PI3K/AKT pathway.