Figure 3: Protein interaction of p53 with Parkin and domain analysis. | Nature Communications

Figure 3: Protein interaction of p53 with Parkin and domain analysis.

From: Cytosolic p53 inhibits Parkin-mediated mitophagy and promotes mitochondrial dysfunction in the mouse heart

Figure 3

(a) Whole-cell lysates of HL-1 cells treated with 0.02 μM DOX and transfected with FLAG-tagged Parkin or the FLAG-empty vector were immunoprecipitated with the anti-Flag-M2 antibody and blotted with anti-p53 and Flag antibodies. (b) Whole-cell lysates of rat neonatal cardiomyocytes treated with the indicated concentrations of H2O2 and DOX for 24 h were immunoprecipitated with the anti-Parkin antibody. (c) Cytosolic heart lysates of adult and aged mice were immunoprecipitated with anti-Parkin, p53 and control IgG antibodies. (d) Schematic representation of N-terminally FLAG-tagged WT Parkin and various mutants of Parkin. (e) The association of endogenous p53 with various Parkin mutants in DOX-treated HeLa cells. The immunoprecipitation assay revealed that the RING0 domain was essential for the interaction with p53. (f) Schematic representation of the GST–p53 fusion proteins. (g) GST pull-down assays using GST–p53 fusion proteins and in vitro transcribed/translated FLAG-Parkin. Residues 81–160 of p53 were sufficient to bind Parkin.

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