Figure 3
The Akt signaling pathway and its downstream effectors were altered by ADQ. SK-Hep1 cells were treated with ADQ for 24 h in 10% FBS-containing media. (A) The expression levels of p-Akt (Ser473), Akt, and α-tubulin were detected by specific antibodies. (B) The expression levels of p53, BAX, and α-tubulin were detected by specific antibodies. (C) SK-Hep1 cells were co-transfected with the pSOD-Luc reporter and gWIZ-GFP. The cells were treated with the indicated concentrations of ADQ for 24 h in culture media containing 10% FBS. The luciferase activities were normalized against the GFP levels. (D) The transcription levels of CDKN1A, CDKN1B, CDKN2B, CDKN2D, and GAPDH were analyzed by RT-PCR. (E) The expression levels of p21 and α-tubulin were detected by specific antibodies. (F) The cells were harvested after 24 h of incubation with ADQ in 10% FBS media and analyzed. The bar graph shows the distribution of cells in the different phases of the cell cycle. (G) SK-Hep1 cells (1500 cells/well) were seeded in 96-well plates. Cells were treated with the indicated concentration of ADQ in media containing 10% FBS. At each indicated time point, cellular growth was measured using the Ez-Cytox solution. Data are representative of three experiments and expressed as the means ± SEM. Data were analyzed by one-way ANOVA followed by Holm-Ĺ Ădák's post hoc test; *p < 0.05, **p < 0.01, and ***p < .001 relative to the ADQ-untreated control.
