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A–C
Luciferase (LUC) activity of the human IGF1R promoter (left) and expression of NRF2 and IGF1Rβ (right) in GSTZ1 knockout (GSTZ1‐KO) HepG2 and GSTZ1‐1‐overexpressing (GSTZ1‐OE) Huh7 cells. GSTZ1‐KO cells were treated with or without brusatol (Bru, 40 nM) for 24 h (A) and transfected with pSEB‐Vector or pSEB‐Keap1 (B) GSTZ1‐OE cells were treated with or without tertiary butylhydroquinone (tBHQ, 40 μM) for 3 h (C).
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D
Co‐immunoprecipitation assay shows NRF2–SP1 interactions in MHCC‐97H cells.
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E
Chromatin immunoprecipitation assay was conducted using extracts of MHCC‐97H cells treated with or without Bru (40 nM) for 24 h (left), and with or without tBHQ (40 μM) for 3 h (right). IgG and anti‐Histone H3 (H3) were used as negative and positive controls, respectively.
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F
Expression of IGF1Rβ and NRF2 in HepG2 cells treated with phenylalanine (Phe, 0, 0.5, 1.0, 2.0, and 4.0 mM, top) and succinylacetone (SA, 0, 50, 100, 200, and 500 μM, bottom) for 48 h, as assessed by Western blotting.
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G
Chromatin immunoprecipitation assay of extracts from MHCC‐97H cells treated with or without Phe (2.0 mM, left) and SA (200 μM, right) for 48 h; IgG and anti‐Histone H3 were described as above.
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H
IGF1Rβ expression in HepG2 cells treated with Phe (2.0 mM, top) and SA (200 μM, bottom) for 48 h, then respectively added 2‐(2‐nitro‐4‐trifluoromethylbenzoyl)‐1,3‐cyclohexanedione (NTBC, 0, 4, 7, 14, 28 μg/ml) for the last 12 h.
‐test (two groups) or one‐way ANOVA followed by the Tukey test (more than two groups). Abbreviations: Vec, vector; ns, not significant.
