NF-κB–dependent transcriptional activation promotes DNA demethylation and results in IL-32 up-regulation after long-term TNFα treatment.
A, schematic representation of a κB site (GGGAGTTTCC) in the IL-32 promoter. B, p65 ChIP-Seq results showed that p65 is enriched at the κB site of the IL-32 promoter after 12 h of TNFα treatment. C, Western blot data validating the RELA KO cell line. D, RT-qPCR results revealed impaired IL-32 induction in RELA KO cells. Averages from three independent experiments are shown, and error bars represent standard deviation. E, locus-specific bisulfite sequencing results showed that the IL-32 CpG island DNA demethylation reaction that occurs during long-term TNFα stimulation is impaired in RELA KO cells. F, RT-qPCR results showed that up-regulation of IL-32 transcription after long-term TNFα treatment is impaired in RELA KO cells. Averages from three independent experiments are shown, and error bars represent standard deviation. d, day.