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. 1996 Dec 24;93(26):15190–15194. doi: 10.1073/pnas.93.26.15190

Figure 1.

Figure 1

Sequestered wt p53 fails to bind to its specific DNA target. (A) EMSA of 5 μg of cytoplasmic extract of LAN-5 cells, using the p53 CON responsive element without (lane 2) or with 50-fold excess of specific (lane 3) and various nonspecific (lanes 4–6) competitor DNA. Only a nonspecific band is seen. Lanes 2–6 contain PAb 421 for maximum stimulation of binding. In contrast, control baculoviral human wt p53 (lanes 12–15) shows a specific band which supershifts and is greatly enhanced in the presence of PAb 421. Nuclear extracts of SH DD1-10 cells (lanes 7–9) fail to show a specific shift, as do nuclear extracts from SaOs 2 p53-null cells (lanes 10 and 11). Lanes 1–10 and 12–15 are from the same gel. See text for further details. (B) EMSA of 5 μg of cytoplasmic extracts of SK-N-SH cells using p53 CON without (lanes 2 and 3) or with 50-fold excess of specific (lane 4) and mutant (lane 5) competitor DNA. As controls, 5 μg of nuclear ML-1 (lanes 11–15) and LAN-5 extracts (lanes 21–24) stimulated with actinomycin D show a specific supershifted band, identical to the one with baculoviral p53. PAb 421 was added as indicated.