Figure 3.

NB124 stabilises the mutant p53 mRNA and restores production of the full-length p53 protein in HDQ-P1 cancer cells. (A) HDQ-P1 cells carrying the endogenous R213X nonsense mutation were treated with NB124 (0.05, 0.1, 0.2, 0.4 and 0.8 mg/mL), G418 (0.05, 0.1 and 0.2 mg/mL) or apramycin (1 mg/mL) for 48 h. Levels of mutant p53 mRNA were determined by RT-qPCR. The results of each experiment are expressed relative to the amount of mRNA in the absence of treatment. Median values are presented, together with the SEM (n = 3). (B) HDQ-P1 cells were treated with gentamicin (0.8 mg/mL), G418 (0.2 mg/mL) or NB124 (0.05, 0.1 and 0.4 mg/mL) for 48 h. Western-blot membranes were probed with the DO-1 antibody directed against the N-terminus of p53 and an anti-actin antibody was used as the loading control. An extract from LoVo cells (p53WT) was used as a control.