Figure 3.

NRF1 targets multiple germ cell–specific genes in testes. A) Luciferase activities driven by Lin28a or Ddx25 promoters containing NRF1 binding sites were examined upon cotransfection of an NRF1 transgene or a vector control (ctrl). Relative firefly luciferase activity was normalized to Renilla activity and represented as means ± 1 sem from 6 biologic replicates. ***P < 0.001. B) NRF1 is mainly enriched at promoter regions of its targeted genes. C) Confirmation of NRF1 target sites by ChIP-PCR with an antibody against NRF1 from wild-type adult testes. D) GC1 cells were treated with or without 5′-Aza and transfected with a vector control or a NRF1-expressing plasmid. Transcript levels of NRF1 target genes were measured by real-time RT-PCR and represented as mean ± 1 sem from 6 biologic replicates. *P < 0.05, **P < 0.01, ***P < 0.001. Minus indicates without; plus denotes in the presence of 5′-Aza or the NRF1 transgene.