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. 2019 Jan 22;20(3):465. doi: 10.3390/ijms20030465

Figure 6.

Figure 6

Vanillic acid inhibits the proliferation of HCT116 cells via blocking cell cycle progression in the G1 phase. (A) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Subsequently, cell cycle status was analyzed by by FACSCalibur flow cytometry. (B) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h. Cyclin D1 and c-Myc were detected by Western blot analysis. Levels of tubulin were used as a loading control. (C) Data were shown as mean ± SD (n = 3). ** p < 0.01, *** p < 0.001, compared with hypoxia control. (D,E) HCT116 cells and SW620 cells were pretreated without or with indicated concentration of vanillic acid (Van), then cultured under normoxic or hypoxic conditions for 12 h, and then the medium was replaced by fresh medium. Cells were allowed to grow for 10 d in normoxic or hypoxic conditions. (F,G) HCT116 cells were pretreated without or with indicated concentration of vanillic acid (30 µM), rapamycin (100 nM), and PD98059 (50 µM), cultured under normoxic or hypoxic conditions, and then the medium was replaced by fresh medium. Cells were allowed to grow for 10 d.