To confirm the role of RA on Nrf2-mediated protection, Nrf2-KO endothelial cells were generated using the CRISPR/Cas9 gene-editing tool. EA.hy926 cells were transfected with Nrf2-CRISPR/Cas9-GFP plasmid (sc-400017, Santa Cruz, USA). GFP+ cell expression was screened using FACS. Western blot analysis for Nrf2-expression was performed to confirm successful Nrf2-depletion. Further, the Nrf2 role in RA which facilitated the protection of endothelial cells against ER stress was assessed using Nrf2 KO cells by assessing the expression of ER stress markers and cell viability.

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