FIG. 5.
DNA binding of ts p53 is not affected by PKR. EMSA was performed with cell extracts from control (CON) zeocin-resistant PKR+/+ and PKR−/− cells (lanes 1 to 3 and 7 to 9) or cells expressing ts p53 (lanes 4 to 6 and 10 to 12) which were incubated at either 37°C (lanes 1, 4, 7, and 10) or 32°C (lanes 2, 3, 5, 6, 8, 9, 11, and 12). Cell extracts (10 μg of protein) were incubated with a 32P-labeled dsDNA oligonucleotide which contains the consensus DNA-binding site of p53. Binding reaction mixtures contained 100 ng of affinity-purified anti-p53 MAb PAb421, which induces the sequence-specific DNA binding of p53. A 200-fold excess of unlabeled dsDNA oligonucleotide was added in cold competition reactions (lanes 3, 6, 9, and 12).