Data in support of sustained upregulation of adaptive redox homeostasis mechanisms caused by KRIT1 loss-of-function

Cinzia Antognelli; Eliana Trapani; Simona Delle Monache; Andrea Perrelli; Claudia Fornelli; Francesca Retta; Paola Cassoni; Vincenzo Nicola Talesa; Saverio Francesco Retta

doi:10.1016/j.dib.2017.12.026

Data in support of sustained upregulation of adaptive redox homeostasis mechanisms caused by KRIT1 loss-of-function

Data Brief. 2017 Dec 13:16:929-938. doi: 10.1016/j.dib.2017.12.026. eCollection 2018 Feb.

Authors

Cinzia Antognelli¹, Eliana Trapani², Simona Delle Monache³, Andrea Perrelli², Claudia Fornelli², Francesca Retta², Paola Cassoni⁴, Vincenzo Nicola Talesa¹, Saverio Francesco Retta²

Affiliations

¹ Department of Experimental Medicine, University of Perugia, Italy.
² Department of Clinical and Biological Sciences, University of Torino, Italy.
³ Department of Biotechnological and Applied Clinical Sciences, University of L'Aquila, Italy.
⁴ Department of Medical Sciences, University of Torino, Italy.

Abstract

This article contains additional data related to the original research article entitled "KRIT1 loss-of-function induces a chronic Nrf2-mediated adaptive homeostasis that sensitizes cells to oxidative stress: implication for Cerebral Cavernous Malformation disease" (Antognelli et al., 2017) [1]. Data were obtained by si-RNA-mediated gene silencing, qRT-PCR, immunoblotting, and immunohistochemistry studies, and enzymatic activity and apoptosis assays. Overall, they support, complement and extend original findings demonstrating that KRIT1 loss-of-function induces a redox-sensitive and JNK-dependent sustained upregulation of the master Nrf2 antioxidant defense pathway and its downstream target Glyoxalase 1 (Glo1), and a drop in intracellular levels of AP-modified Hsp70 and Hsp27 proteins, leading to a chronic adaptive redox homeostasis that sensitizes cells to oxidative DNA damage and apoptosis. In particular, immunoblotting analyses of Nrf2, Glo1, AP-modified Hsp70 and Hsp27 proteins, HO-1, phospho-c-Jun, phospho-ERK5, and KLF4 expression levels were performed both in KRIT1-knockout MEF cells and in KRIT1-silenced human brain microvascular endothelial cells (hBMEC) treated with the antioxidant Tiron, and compared with control cells. Moreover, immunohistochemistry analysis of Nrf2, Glo1, phospho-JNK, and KLF4 was performed on histological samples of human CCM lesions. Finally, the role of Glo1 in the downregulation of AP-modified Hsp70 and Hsp27 proteins, and the increase in apoptosis susceptibility associated with KRIT1 loss-of-function was addressed by si-RNA-mediated Glo1 gene silencing in KRIT1-knockout MEF cells.

Keywords: Adaptive redox homeostasis; Antioxidant defense; Argpyrimidine-modified heat-shock proteins; CCM1/KRIT1; Cerebral cavernous malformations; Cerebrovascular disease; Glyoxalase 1 (Glo1); Heme oxygenase-1 (HO-1); Nuclear factor erythroid 2-related factor 2 (Nrf2); Oxidative DNA damage and apoptosis; Oxidative stress; Redox signaling; c-Jun.