MDM2 and HSP70 colocalize with p53R175H in an aggressome.

<p>(<b>A</b>) Immunostaining of MEF cells (<i>Trp53^−/−, Mdm2^−/−</i>) transfected with plasmids encoding p53 R175H, HSP70 and optionally MDM2 (lower panel) shows that only in the presence of MDM2, A11 antibodies (labelling amyloid-oligomers) recognize aggregates composed of mutant p53 and HSP70. (<b>B</b>) Immunostaining of MEF cells (<i>Trp53^−/−, Mdm2^−/−</i>) transfected with plasmids encoding p53R175H, MDM2 and HSP70 revealed recruitment of both <b>(1)</b> the E3 ubiquitin ligase and <b>(2)</b> the chaperone to the large inclusion body. (<b>3)</b> The inclusion is formed at the centrosome marked by the presence of gamma-tubulin, suggesting that the large inclusion body containing p53 R175H is an aggresome. (<b>4</b>) Disruption of microtubules with nocodazole treatment impairs protein transport and results in the disappearance of the aggresome; smaller, scattered cytoplasmic p53-containing aggregates form instead. Scale bar 10 µm.