H₂O₂/DEM induced the activation of the Keap1/Nrf2 signaling pathway and the transcription of target genes. The changes in the relative mRNA expression of (A) keap1a and keap1b, (B) sMaf family genes, (C) nrf2a and nrf2b, (D) antioxidation-related genes were analyzed before and after treatment. n = 30, ** p < 0.01, * p < 0.05, ns p > 0.05, mean ± SEM.

Methylation analysis of promoter regions of the Keap1 /Nrf2 signaling pathway-related genes before and after treatment. (A) Methylation prediction of promoter regions of genes related to the Keap1 /Nrf2 signaling pathway, Methylation analysis of promoter regions of (B) keap1 and nrf2, (C) Mafg2, Mafk, and Maft. Each cycle is a CpG in the DNA sequence represented by either a closed circle (methylated cytosine) or an open circle (unmethylated cytosines). Each column of CpGs represents one colony sequenced, with a total of 10 positive clones.

The level of methylation of maft promoter after inhibitors treatment. Each cycle is a CpG in the DNA sequence represented by either a closed circle (methylated cytosine) or an open circle (unmethylated cytosines). Each row of CpGs represents one colony sequenced, with a total of 10 positive clones.

The expressions of the Nrf2/ARE signaling pathway-related genes and target genes under different stimuli after the addition of decitabine and azacytidine (A) keap1a, (B) keap1b, (C) nrf2a, (D) nrf2b, (E) maft, (F) gclc, (G) gstp1, (H) nqo-1, (I) sod1. n = 30, *** p < 0.001, ** p < 0.01, * p < 0.05, ns p > 0.05, mean ± SEM.

Analysis of methyltransferase expression (A) after H₂O₂/DEM treatment, after methyltransferase inhibitors treatment, analysis of the expression of (B) dnmt1, (C) dnmt3a1, (D) dnmt3a2, (E) dnmt3b1, (F) dnmt3b2, (G) dnmt3b3, (H) dnmt3b4. n = 30, *** p < 0.001, ** p < 0.01, * p < 0.05, ns p > 0.05, mean ± SEM.