GSTZ1‐1 Deficiency Activates NRF2/IGF1R Axis in HCC via Accumulation of Oncometabolite Succinylacetone AWestern blotting shows endogenous GSTZ1‐1 protein.

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GSTZ1‐1 Deficiency Activates NRF2/IGF1R Axis in HCC via Accumulation of Oncometabolite Succinylacetone AWestern blotting shows endogenous GSTZ1‐1 protein expression in HCC cell lines and MIHA cells. BOverexpression of GSTZ1‐1 in Huh7 (left) and SK‐Hep1 (middle) cells and knockout of GSTZ1 in HepG2 (right) cells were confirmed by immunoblot assay. The GSTZ1‐1‐overexpressing (GSTZ1‐OE) cell model was established by infecting hepatoma cells with adenoviruses expressing GSTZ1‐1 (AdGSTZ1). Adenoviruses expressing green fluorescent protein (AdGFP) were used as a control. The GSTZ1 knockout (GSTZ1‐KO) cell model was established using the CRISPR/Cas9 system. C–EProliferation ability of GSTZ1‐OE Huh7 and SK‐Hep1 cells, and GSTZ1‐KO HepG2 cells. (C) Representative images (left) and quantification (right) of EdU‐positive cells. Scale bars: 50 μm. (D) Cell growth curves. (E) Representative images (left) and quantification (right) of colony formation capacity. Values are shown as means ± SD (n = 3), *P < 0.05, **P < 0.01, ***P < 0.001, Student's t‐test (two groups) or one‐way ANOVA followed by the Tukey test (more than two groups). Abbreviations: DAPI, 4′,6‐diamidino‐2‐phenylindole; EdU, 5‐ethynyl‐2′‐deoxyuridine. Source data are available online for this figure. IF THIS IMAGE HAS BEEN PROVIDED BY OR IS OWNED BY A THIRD PARTY, AS INDICATED IN THE CAPTION LINE, THEN FURTHER PERMISSION MAY BE NEEDED BEFORE ANY FURTHER USE. PLEASE CONTACT WILEY'S PERMISSIONS DEPARTMENT ON PERMISSIONS@WILEY.COM OR USE THE RIGHTSLINK SERVICE BY CLICKING ON THE 'REQUEST PERMISSIONS' LINK ACCOMPANYING THIS ARTICLE. WILEY OR AUTHOR OWNED IMAGES MAY BE USED FOR NON-COMMERCIAL PURPOSES, SUBJECT TO PROPER CITATION OF THE ARTICLE, AUTHOR, AND PUBLISHER. EMBO J, Volume: 38, Issue: 15, First published: 28 June 2019, DOI: (10.15252/embj.2019101964)