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. 2020 May 21;11:2549. doi: 10.1038/s41467-020-16312-7

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© The Author(s) 2020

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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Fig. 8 — a Cell survival by neutral red assay in WT and PGAM5^−/− ARPE-19 cells after overnight NaIO₃ treatment using indicated concentrations. n = 4, *p < 0.05, ****p < 0.0001, two-way ANOVA Tukey’s multiple comparisons test. Error bars, mean ± s.d. b Quantification of PI staining showing cell death in WT and PGAM5^−/− ARPE-19 cells after overnight NaIO₃ (10 mM) treatment using Nikon Elements software and ImageJ. n = 4, ***p = 0.0002, two-tailed unpaired t-tests, error bars, mean ± s.d. c Representative PI staining in b. Scale bar = 250 µm. n = 4. d Upregulation of anti-oxidative genes (NRF2, FOXO4 and SOD1) in PGAM5^−/− ARPE-19 cells compared to control cells after short-term (1 week) culture. Data were normalized to GAPDH. n = 3, ***p = 0.001(Nrf2), p = 0.0007(Foxo4), p = 0.0006, two-tailed unpaired t-tests, error bars, mean ± s.d. e Downregulation of anti-oxidative genes (NRF2, FOXO4 and SOD1) in PGAM5^−/− ARPE-19 cells compared to control cells after long-term (8 weeks) culture. Data were normalized to GAPDH. n = 4, ***p = 0.0003; **p = 0.0022; *p = 0.0258, two-tailed unpaired t-tests, error bars, mean ± s.d. f Upregulation of NRF2 protein in PGAM5^−/− ARPE-19 cells compared to control cells after short-term (1 week) culture. Data were normalized to β-actin. n = 4. g Quantification of data in f. n = 4, *p = 0.042, two-tailed unpaired t-tests, error bars, mean ± s.d. h Downregulation of NRF2 protein in PGAM5^−/− ARPE-19 cells compared to control cells after long-term (8 weeks) culture. Data were normalized to β-actin. n = 5. i Quantification of data in h. n = 5, ***p = 0.0003, two-tailed unpaired t-tests, error bars, mean ± s.d. j Nrf2 expression as measured by qRT-PCR in the RPE/choroid of WT and Pgam5^−/− mice at 2 and 18-months old. Data were normalized to Gapdh. n = 3, ****p < 0.0001, two-way ANOVA Tukey’s multiple comparisons test. Error bars, mean ± s.d. For assays in the figure, n represents the number of biologically independent experiments unless otherwise specified. Images were captured under same settings, and representative images were shown. Source data are available as a Source Data file.