De novo aggregation of soluble p53 with recombinant p53 aggregates.

<p>(A) NIH3T3 cells were transfected with p53GFP or pEGFPN1 24 h prior to addition of 1 µM purified p53 protein incubated at 4 °C or 42 °C during 24 h. (B) % of cells containing p53GFP aggregates when incubated with either 1 µM p53 aggregates, Aβ oligomers or without aggregates. Mean value of 3 experiments is shown (*, p < 0,001); 100 cells counted per experiment. (C) Purified p53C protein was resolved by 10% SDS-PAGE and stained with Coomassie Blue (lane 1) or analyzed by Western Blot (lane 2). Transmission electron microscopy showing aggregated p53C. (D) NIH3T3 cells were incubated with 1 µM DyLight 650-conjugated p53C aggregates for 24 h, treated with 0,25% Trypsin/EDTA and re-plated onto microscope slides that were incubated with 100 µM etoposide for 2 h before visualization by confocal microscopy. Scale bars: 5 µm (A), 500 nm (B) and 10 µm (C).