Skip to main content
. Author manuscript; available in PMC: 2016 Oct 18.
Published in final edited form as: J Invest Dermatol. 2016 May 7;136(8):1656–1663. doi: 10.1016/j.jid.2016.04.022

Figure 5. The Lce1e gene is a direct downstream target of NRF2.

Figure 5

(a) Chromatin immunoprecipitation assays were performed on nuclear extracts isolated from LKO and WT back skin using the anti-NRF2 antibody. LKO skin showed enrichment of the region flanking AREs, which are upstream of Sprr2d and Lce1e genes (WT n = 3, LKO n = 3, **P < 0.01, ***P < 0.005, ****P < 0.001). (b) Schematic representation of putative promoter regions of the Sprr2d and Lce1e genes (left panel). The Sprr2d gene has an AREs at −780 bp and −3.6 kbp upstream, and Lce1e has an ARE at −3.7 kbp upstream from the transcription start site. Luciferase expression constructs containing the promoter regions of the Sprr2d and Lce1e genes show increased activity in a tBHQ dose-dependent manner (right panel). (c) Mutations in the ARE reduce Lce1e promoter activity. Two different mutations were introduced into the ARE in the promoter region of the Lce1e gene and inserted into the luciferase expression vector. Constructs containing both mutations exhibit reduced activity in response to tBHQ compared with the expression construct containing the WT Lce1e ARE (WT n = 6, Mutants [Muts] n = 6, **P < 0.01). ARE, antioxidant response element; LCE, late cornified envelope; LKO, loricrin knockout; NRF2, NF-E2-related factor 2; tBHQ, tert-butylhydroquinone; WT, wild type.