Fig. 5.

Effect of XWL-1-48 on PI3K/Akt/Mdm2 pathway. a-c MCF-7 cells were treated by XWL-1-48 (1, 3, 10 μM) for 24 h, protein expression of p-Akt, Akt, p-Mdm2 and Mdm2 were measured by Western blot analysis. The changes of p-Akt/Akt and p-ATM/ATM were quantified using Image J software; d-f MCF-7 cells were exposure to a known pan-PI3K inhibitor LY294002 or XWL-1-48 for 24 h, protein expression level of p-Akt, Akt, p-Mdm2 and Mdm2 was determined using immunoblot; g After treated with XWL-1-48 (10 μM) for 0, 2, 4, 6, 12, 24 h, cells were harvested. The protein expression level of Mdm2 was analyzed by western blot; h After incubated with XWL-1-48 (1, 3, 10 μM) for 24 h, the mRNA expression of Mdm2 was detected by RT-PCR; i MCF-7 cells were exposed to 100 μg/ml cycloheximide (CHX, known as protein synthesis inhibitor) with or without XWL-1-48 (10 μM) for 0, 10, 30, 60, 90, 120, 180 and 240 min to block protein synthesis. The cells were collected for Western blot analysis. A representative result is shown from at least three independent experiments. *p < 0.05, **p < 0.01 vs. control, respectively