Fig. 5

NRF2 is required for nuclear LDHA-induced antioxidant responses. a E7 induces the expression of antioxidant and Wnt target genes. qPCR detecting antioxidant and Wnt target genes in vector (treated with or without H₂O₂) or HPV16 E7-expressing HT-3 cells treated with or without NAC as indicated. b E7 enhances the H3K79 dimethylation level at gene bodies. ChIP-qPCR showing the percentage of H3K79me2 enrichment at SOD1, CAT, CTNNB1, and MYC gene body relative to input genomic DNA in vector or HPV16 E7-expressing HaCaT cells treated with or without NAC. c EPZ004777 blocks increased NQO1 and GCLC gene expression induced by E7 and H₂O₂. qPCR detecting NQO1 and GCLC genes in vector (treated with or without H₂O₂) or HPV16 E7-expressing HaCaT cells treated with or without NAC and 3 μM EPZ004777 for 24 h as indicated. d, e NRF2 KO or NRF2 inhibition blocks increased NQO1 and GCLC gene expression induced by E7 and H₂O₂. qPCR detecting NQO1 and GCLC genes in HeLa NRF2 KO cells treated with or without H₂O₂ (e), and vector or HPV16 E7-expressing HaCaT cells treated with or without 10 μM ML385 for 24 h (f) as indicated. f LDHA KO decreases NQO1 and GCLC gene expression activated by H₂O₂. qPCR detecting NQO1 and GCLC genes in HeLa LDHA KO cells upon H₂O₂ coupled with or without extended NAC treatment as indicated. g LDHA KO attenuates the H3K79 dimethylation level at gene bodies. ChIP-qPCR showing the percentage of H3K79me2 enrichment at SOD1, CAT, CTNNB1, and MYC gene body relative to input genomic DNA in HeLa LDHA KO and NRF2 KO cells treated with or without H₂O₂. For ChIP-qPCR assay, rabbit IgG was included as a negative control. H₂O₂ was used as 10 μM for 6 h, and NAC was used as 1 mM for 6 h. Results are representative of three independent experiments. All data are shown as mean ± SEM. The p values were determined by two-tailed t-test. The values of p < 0.05 were considered statistically significant. *, **, and *** denote p < 0.05, p < 0.01, and p < 0.001, respectively. NS means non significant