Fig 2: STRING network of p53 knockdown HaCaT proteins with significant regulation levels. Expression ratios were mapped to the nodes using blue-red gradient, where blue is related to the downregulated and red to the upregulated proteins. Proteins without interaction partners were omitted from the visualization. Network edges represent the confidence of interaction. The required interaction score was set to >0.7.

Fig 3: Effect of CHCHD4 reduction on p53 expression. (a) Immunoblotting of p53 in the homogenate (top panel, Homo-p53), cytosolic (Cyto-p53), mitochondrial (Mito-p53), and nuclear fraction (bottom panel, Nucl-p53) of normal control (Cont) mice and at 6 or 24 h after HI. The p53 band was decreased in both mitochondrial and nuclear fractions, and was more pronounced at 24 h after HI in both the Wt and Het groups. (b) Bar graphs show the quantification of p53 immunoblotting in the mitochondrial fraction. (c) Bar graphs show the quantification of p53 immunoblotting in the nuclear fraction of the Wt and Het groups (n=6 per group). (d) ELISA assay of total and phosphorylated p53 in the mitochondrial fraction of controls and at 6 h after HI in both Wt and Het mice. (e) Total and phosphorylated p53 in the cytosolic fraction of controls and at 6 h after HI in both Wt and Het mice (n=5 per group). (f) The mRNA expression levels of p53 pathway-related genes, including p53, p21, Peg3, PUMA, Usp7, and Mdm2, were quantified by RT-qPCR both in Wt and Het mice, and normalized with the reference gene Ywhaz at different time points after HI. n=6 for Wt control group, n=5 for Het control group, n=6 for both the Wt and Het groups at 6 h after HI, and n=6 for Wt and n=5 for Het at 24 h after HI. *P<0.05, **P<0.01, ***P<0.001