Figure 8. Reactivation of CDKN1A (p21) tumour suppressor on mRNA and protein level by DAC, AZA and vitamin C in colon cancer cells.
HCT116 cells were incubated with increasing concentrations of DAC or AZA, alone or in combination with 0 μM (striped bars), 10 μM (grey bars) and 50 μM (black bars) vitamin C for 72 h. The mRNA levels of CDKN1A (p21) after treatment with DAC (A) and AZA (B) were then calculated by 2-ΔΔCT method in relation to HMBS, which served as a control. Additionally, the protein levels of CDKN1A (p21) were determined by Western Blotting and ECL detection (C). The blotting intensities were normalized to β-actin and the untreated control. The experiments were conducted in triplicate. Statistical significance of the treated groups to the untreated control was calculated using 2-way ANOVA and Tukey post-test: *p < 0.05, **p < 0.01, error bars = SD; n=3.