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. 2019 Feb 4;10(2):96. doi: 10.1038/s41419-019-1316-8

Fig. 4. Pin1 expression was increased by heat stress (HS) and required for efficient mitochondrial localization of p53.

Fig. 4

The p53+/+ mouse aortic endothelial cells (MAECs) were subjected to intense HS (43 °C) for 2 h and then further incubated at 37 °C for 0, 3, 6, or 9 h (a–b). a Pin1 and cyclin D1 protein levels were assessed by western blot. b Quantification of Pin1 activity at the indicated time points after HS. c Quantification of Pin1 mRNA at the indicated time points after HS. The p53-/- and p53+/+ MAECs were exposed to 43 °C for 2 h and then further incubated at 37 °C for 6 h (d–f). d Pin1 and p53 protein levels were assessed by western blot. e Quantification of Pin1 activity induced by HS. f Quantification of Pin1 mRNA induced by HS. Mice were put through the same HS assay as previously described. The mice were sacrificed at 0, 6, and 9 h after exposure to HS and the aortic endothelium was isolated (g–i). g Pin1 protein levels were measured by western blot. h Quantification of Pin1 activity at the indicated time points after HS. i Quantification of Pin1 mRNA at the indicated time points after HS. Mice were put through the same HS assay as previously described. The mice were sacrificed at 6 h after exposure to HS and the aortic endothelium was isolated (j–l). j Pin1 protein levels were measured by western blot. k Quantification of Pin1 activity induced by HS. l Quantification of Pin1 mRNA induced by HS. Human umbilical vein endothelial cells (HUVECs) were exposed to intense heat (43 °C) for 2 h and then incubated at 37 °C for 6 h (m–o). m Expression of P-Ser 46 p53 and p53 were determined by western blot. n Quantification of the P-Ser 46 p53 and p53 ratio. o The p53+/+ MAECs were exposed to 43 °C for 2 h and were further incubated at 37 °C for 6 h. Cell lysates were normalized for p53 protein levels and then subjected to co-immunoprecipitation to analyze interactions between endogenous p53 and Pin1. IgG served as the negative control. *P < 0.05 compared with the control group (37 °C) in p53+/+ MAECs or the control group in wild-type mice, n = 6