Aggregation propensity of p53 is conformation-dependent and promoted by HSP70.
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(A) Unfolding of p53 induces its nuclear aggregation. H1299 cells were transfected with plasmids encoding WT p53 (1, 3, 5) or p53 V143A (2, 4) and the next day incubated for 24 hours at the indicated temperature. In case of 42°C treatment, cells were incubated for 23 hours at 37°C and for 1 hour at 42°C. Small nuclear p53-containing aggregates can be seen in 4 and 5. Scale bar 10 µm. (B) HSP70 delays the disappearance of severely unfolded p53 temperature sensitive mutant V143A. Lysates from H1299 cells, transfected with plasmids encoding for indicated proteins, were subjected to p53 conformation-specific immunoprecipitation and analysed by western blotting. For WT conformation two exposures of the same blot are shown.