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. 2020 Apr 6;16(11):1901–1916. doi: 10.7150/ijbs.44343

Figure 6.

Figure 6

Effect of MHY2245 on Akt/mTOR pathways in SKOV3 cells. (A) Cells were treated with vehicle control, MHY2245 (0.03, 0.1 or 0.3 µM), or doxorubicin (DOX; 1 µM) for 48 h, and then the expression of PI3K, p-PI3K, Akt, p-Akt, PTEN, p-PTEN, mTOR, and p-mTOR levels were analyzed by Western blot. β-Actin was used as a loading control. (B) Representative histogram showing the expression levels were quantified using Image J software compared to β-actin. Data expressed as mean ± SD of triplicate experiments. *p < 0.05 and **p < 0.01 and ***p < 0.01versus the control group. (C) Cells were treated with vehicle control, MHY2245 (0.03, 0.1 or 0.3 µM) or DOX (1 µM) for 48 h and the acetylated p53 and p53 levels were analyzed by Western blot. β-Actin was used as a loading control (upper). SKOV3 cells were treated with MHY2245 or DOX for 48 h and fixed for immunostaining with antibodies against acetylated p53. Immunofluorescence and DAPI images were taken at a 50× magnification and merged (Lower). (D) Cells were treated with vehicle control, MHY2245 (0.03, 0.1 or 0.3 µM) or DOX (1 µM) for 48 h and the MDM2 and p-MDM2 expression was analyzed by Western blot. β-Actin was used as a loading control (upper). SKOV3 cells were treated with MHY2245 or DOX for 48 h and fixed for immunostaining with antibodies against p-MDM2. Immunofluorescence images were taken with a 50× magnification (lower). Scale bar, 10 µm.