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. 2022 Dec 30;132(2):208–222. doi: 10.1161/CIRCRESAHA.122.321153

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© 2023 The Authors.

Circulation Research is published on behalf of the American Heart Association, Inc., by Wolters Kluwer Health, Inc. This is an open access article under the terms of the Creative Commons Attribution Non-Commercial-NoDerivs License, which permits use, distribution, and reproduction in any medium, provided that the original work is properly cited, the use is noncommercial, and no modifications or adaptations are made.

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Figure 2. — The nuclear translocation of ASPP1 and p53 are interdependent. A and B, Effects of ASPP1 overexpression or knockdown on the total protein level and nuclear distribution of p53 in neonatal mouse ventricular cardiomyocytes (NMVCs) by Western blot; n=5 for total p53 level (Kruskal-Wallis, followed by false discovery rate [FDR] method of Benjamini and Hochberg test); n=6 for nuclear and cytoplasmic p53 levels (1-way ANOVA, followed by Tukey post hoc multicomparisons test); ns, not significant. C and D, Immunofluorescent staining was performed to analyze the effects of ASPP1 overexpression or knockdown on p53 nuclear translocation in NMVCs (1-way ANOVA, followed by Tukey post hoc multicomparisons test); n=6; ns, not significant. Scale bar=20 μm. E, Effects of p53 knockdown on ASPP1 nuclear translocation in NMVCs (Mann-Whitney U test); n=5. Scale bar=20 μm. F, Effects of importin-β1 knockdown on ASPP1 and p53 nuclear translocation in NMVCs (Mann-Whitney U test); n=5. Scale bar=20 μm. G and H, Effects of ASPP1 overexpression (G) on mRNA levels of Bax (n=9), PUMA (n=9), and Noxa (n=9) or ASPP1 knockdown (H) on mRNA levels of Bax (n=8), PUMA (n=9), and Noxa (n=8) by quantitative real-time PCR (qRT-PCR, 1-way ANOVA, followed by Tukey post hoc multicomparisons test). ASPP1 indicates apoptosis stimulating of p53 protein 1; Bax, Bcl2-associated X; DAPI, 4′,6-diamidino-2-phenylindole; H/R, hypoxia/reoxygenation; and PUMA, p53 upregulated modulator of apoptosis.