Fig 2: The expression level of p53 was assessed using the p53 N-terminal antibody (DO-1) following treatment with either DMSO or PTC124 in four different p53 nonsense mutation clones. It was observed that PTC124 (50 µM) was able to induce re-expression of p53 as red color in H1299-R306X and H1299-R342X clones but not in H1299-W91X and H1299-S94X clones. Hoechst 33342 was used to label the nucleus as blue color.

Fig 3: The expression level of p53 was assessed using the p53 C-terminal antibody (PAb 122) in the presence of either DMSO or PTC124 treatment across four different p53 nonsense mutation clones. Our results showed that PTC124 (50 µM) induced re-expression of p53 as red color in H1299-R306X and H1299-R342X clones but not in H1299-W91X and H1299-S94X clones. Nuclear staining was performed using Hoechst 33342 as the blue color.

Fig 4: The inverse PCR products of p53 nonsense mutation clones. Each number represents each band. Band 1: Kb marker (10: 10 Kb; 8: 8 Kb; 5: 5 Kb; 1: 1 Kb). Bands 2–5: PCR products of W91X, S94X, R306X, and R342X of p53 clones were amplified using CloneAmp HiFi PCR Premix. Bands 6–9: PCR products of W91X, S94X, R306X, and R342X of p53 clones were amplified using Phusion Plus PCR Master Mix. Bands 10–13: PCR products of W91X, S94X, R306X, and R342X of p53 clones were amplified using Phusion High-Fidelity PCR Master Mix.

Fig 5: The sequencing of p53 nonsense mutation clones. The sequencing results of a selected single colony from bands 2–5 of Figure 2. (A) W91X (TGG to TGA, the area enclosed in the red rectangle is the changed A base), (B) S94X (TCA to TGA, the area enclosed in the red rectangle is the changed G base), (C) R306X (CGA to TGA [reversed sequence is TCA], the area enclosed in the red rectangle is the changed A base), and (D) R342X (CGA to TGA [reversed sequence is TCA], the area enclosed in the red rectangle is the changed A base).