Supplemental Data

Files in this Data Supplement:

• Supplementary Figure 1 - (SupplementalFig1.ppt; 445 KB) Time course of p53 Ser20 phosphorylation in normal and WS diploid fibroblasts after H₂O₂ treatment. Normal and WS cells were incubated with either 500 μM H₂O₂ (lanes 2–4 and 6–8) or medium alone (lanes 1 and 5) for 45 min at 37°C. The cells were then collected either immediately (0.75 h, lanes 2 and 6), or 5 h (lanes 3 and 7) and 24 h (lanes 4 and 8). Total cell lysates from the same number of cells were analyzed by immunoblotting with anti-phospho-p53 Ser20, anti-p53, and anti-lamin B antibodies. Similar results were obtained when using MRC-5 and WI-38 cells (normal cells), and AG00780H and AG03141C cells (WS cells).
• Supplementary Figure 2 - (SupplementalFig2.ppt; 204 KB) Time course of p53 and p21^WAF1 accumulation. Normal and WS cells were incubated with either 500 μM H₂O₂ (lanes 2–5 and 7–10) or medium alone (lanes 1 and 6) for 45 min at 37°C. The cells were then collected either immediately (0.75 h, lanes 2 and 7), or 5 h (lanes 3 and 8), 24 h (lanes 4 and 9) and 10 days (10 d) (lanes 5 and 10) after recovery time (40 h). Total cell lysates from the same number of cells were analyzed by immunoblotting with anti-p53, anti-p21, and anti-lamin B antibodies. Similar results were obtained when using MRC-5 and WI-38 cells (normal cells), and AG00780H and AG03141C cells (WS cells).
• Supplementary Figure 3 - (SupplementalFig3.ppt; 37 KB) Number of nucleoli in control and H₂O₂-treated cells. Normal and WS diploid fibroblasts were incubated with either 500 μM H₂O₂ (black bars) or medium alone (white bars) for 45 min at 37°C. The cells were then washed and incubated in complete medium for 7–10 days at 37°C. The nucleoli were visualized in both living and fixed cells, obtaining similar results. The number of cells scored is indicated in the figure. Similar results were obtained when using MRC-5 and WI-38 cells (normal cells), and AG00780H and AG03141C cells (WS cells).