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. 2019 Oct 4;9:14354. doi: 10.1038/s41598-019-50877-8

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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TFEB increases the transcriptional activity and the nuclear localization of Nrf2. (a) The gene expression levels in H4 cells (Control) and H4 cells stably expressing TFEB (TFEB) were analyzed using cDNA microarray as described in Methods. Heat map shows the expression levels for Nrf2-response genes. Shades of red represent up-regulation, and shades of green represent down-regulation. (b) H4 cells (Control) and H4 cells stably expressing TFEB (TFEB) were transiently transfected with the ARE-Luc reporter plasmid and assayed for the luciferase activity. (c) H4 cells stably expressing TFEB fixed with 4% paraformaldehyde were immunostained using anti-TFEB and Nrf2 antibodies. Fluorescence signals were observed using a fluorescence microscope. (d) Nuclear (NF) and cytosolic (CF) fractions from control and TFEB cells were prepared as described in Methods. The cellular locations of TFEB and Nrf2 were analyzed by immunoblotting using anti-TFEB and Nrf2 antibodies, respectively. To examine the purity of the fractionations, the blot was probed with antibodies to lamin (A/C), a marker for nuclear fraction, and tubulin, a marker for cytosolic fraction. Full blots are provided in Supplementary Fig. S10. Data shown are mean ± S.E. of four independent experiments and were analyzed using Student’s t test. (**p < 0.01).