Skip to main content
. 2014 Jan 11;5(3):693–703. doi: 10.18632/oncotarget.1328

Figure 3. Effects of kinase inhibitors on actinomycin D (ActD)-induced p53 expression.

Figure 3

(A) Cells were pretreated with PI3K inhibitors (LY294002 (10 μM) or wortmannin (10 μM)), and an AKT inhibitor (deguelin) for 1 h, followed by treatment with ActD for 24 h in 293 and 293T cells, and for 6 h in HepG2 and Hepa-1c1c7 cells. (B) Cells were pretreated with deguelin, (0.01, 0.02, 0.05, 0.1, and 0.2 μM) for 1 h, followed by treatment with ActD for 24 h in 293 and 293T cells, and for 6 h in HepG2 and Hepa-1c1c7 cells. (C) The MEK1/2 inhibitor (PD98059, 10 μM), JNK inhibitor (SP600125, 50 μM), and p38 inhibitor (SB203580, 25 μM) were applied. Cells were pretreated with kinase inhibitors for 1 h, followed by treatment with ActD for 24 h in 293 and 293T cells, and for 6 h in HepG2 and Hepa-1c1c7 cells. (D) Cells were treated individually with ActD (10 nM), LY294002 (10 μM), wortmannin (10 μM), deguelin (0.2 μM for 293, HepG2, and Hepa-1c1c7 cells, and 0.1 μM for 293T cells), PD98059 (10 μM), SP600125 (50 μM), and SB203580 (25 μM) for 24 h in 293 and 293T cells, and for 6 h in HepG2 and Hepa-1c1c7 cells. The cells were then harvested and cell lysates were analyzed by Western blotting using antibodies against p53, phospho-p53 (Ser15), GAPDH, and β-actin.