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. 2020 Aug 16;12(8):2305. doi: 10.3390/cancers12082305

Figure 3.

Figure 3

Nrf2 is also activated in the absence of an oxidative environment. (A) Nqo1 (left) and G6pd (right) mRNA expression level in surrounding tissue and microdissected GSTP+ preneoplastic nodules. mRNA expression levels were assessed by qRT-PCR. Relative mRNA expression was calculated by using the 2-∆∆CT method and Gapdh as endogenous control. Gene expression is reported as fold-change relative to respective surrounding tissue. Each bar represents mean ± standard error (SEM) of 5 to 21 samples per group. Student’s t-test was used for the evaluation of statistical significance. * p < 0.05; *** p < 0.001; (B) Pie chart representing the percentage (58%; 11/19 nodules) of Nrf2 mutations in GSTP+ preneoplastic lesions; (C) List of Nrf2 mutations identified in GSTP+ nodules. All mutations are confined within the DLG (73%) or ETGE (27%) motif of exon 2 of the Nrf2 gene. V32E is the only mutation at the weak bond (DLG) for Keap1, whereas T80A is the most frequent mutation in the strong domain for Keap1. Asterisks indicate mutations present at both 4 and 6 months; (D) Nqo1 and G6pd mRNA levels in mutated and non-mutated (WT) preneoplastic nodules 6 months after DENA treatment; (E) Comparison of Nqo1 and G6pd mRNA levels between DLG and ETGE motif mutations. mRNA expression levels were assessed by qRT-PCR. Relative mRNA expression was calculated by using the 2-∆∆CT method and Gapdh as endogenous control. Gene expression is reported as fold-change relative to surrounding tissue. Each bar represents mean ± standard error (SEM) of 4 to 17 samples per group. ANOVA with Tukey post-hoc test was used for the evaluation of statistical significance. *p < 0.05; ** p < 0.01; ns, not significant.