Fig. 4

Sevo decreases HES1 expression by orchestrating HES1 promoter methylation via EZH2. A HES1 expression in the cortical tissue of sham-operated, TBI, or Sevo-treated TBI rats measured by RT-qPCR and Western blot analysis. B JASPAR database predicting binding sites of EZH2 to HES1 promoters. C Dual luciferase reporter assay validating the interaction between HES1 promoter and EZH2. D ChIP assay for EZH2 binding to HES1 promoter in the hippocampal neuron. E RT-qPCR and Western blot analysis of HES1 expression in hippocampal neurons after overexpression or silencing of EZH2. F HES1 promoter methylation in hippocampal neurons assessed by MSP. G Expression of HES1 in hippocampal neurons treated with Sevo or 5-Aza-dC determined by RT-qPCR. H Expression of HES1 in hippocampal neurons after treatment with Sevo or 5-Aza-dC detected by RT-qPCR and Western blot analysis. *p < 0.05 vs. sham-operated rats, oe NC-treated 293T cells, cell lysate incubated with IgG antibody, or hippocampal neurons treated with oe NC, Sevo, or DMSO. ^#p < 0.05 vs. TBI rats or hippocampal neurons treated with sh NC. Cell experiments were conducted three times independently