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. 2021 Feb 24;11:140. doi: 10.1038/s41398-021-01261-6

Fig. 2. Nrf2 acts as transcription factor for BDNF.

Fig. 2

A–E Nrf2 acted as a transcription factor for BDNF promoters. A, B BDNF exons I, II, and IV luciferase promoters and SFN or Nrf2 or siRNA-Nrf2 plasmids were treated into HEK293 cells (mean ± SEM, n = 4 per group, one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, and # # #p < 0.001). C, D Results obtained using the luciferase plasmids containing mutation (Mut) at the Nrf2 binding motif of BDNF exon I promoter was compared with the wild type of promoters, respectively (mean ± SEM, n = 4 per group, one-way ANOVA, *p < 0.05, **p < 0.01, and ***p < 0.001). E ChIP-PCR assays demonstrated Nrf2 specifically bound to genomic DNA of BDNF exon I promoter binding motifs. The Nrf2 protein–DNA crosslinking samples were obtained from the HEK293 cells treated with SFN or Nrf2 plasmid or not (control) via co-immunoprecipitating with anti-Nrf2 antibodies. PCR was carried out by using primer pairs at BDNF exon I promoter. PCR assay also included each input sample. The positive control was demonstrated with anti-Histone H3 antibody coupling with GAPDH primers. F The immunofluorescence for Nrf2 and MeCP2. The SFN treated for Neuro-2a cell 24 h. The immunofluorescence was performed for Nrf2 and MeCP2. Scale bar 50 μm.