Figure 6.
Plagl1 regulates the thrombopoietin receptor (Mpl) and neuronal pentraxin 1 (Nptx1) genes by direct transactivation of their respective promoters. (A) ChIP-seq and ChIP-qPCR data were mapped to the Mpl locus. ‘IP reads’ designate reads from the anti-Plagl1 immuno-precipitated fraction. (B) Fragments of the SV40 early (SV40-E) or Mpl (Mpl 0.8 kb, Mpl 1.8 kb) promoters were subcloned into the pGL3 reporter plasmid encoding the Firefly Luciferase, and transfected into Neuro-2a cells. The pRK7-βGal plasmid containing the β-galactosidase cDNA under the control of a CMV promoter was co-transfected. The luciferase activity was normalized to the β-galactosidase activity of transfected cells. Data are displayed as ratio of the normalized luciferase activity in pGL3promoter- to pGL3basic-transfected cells. Plagl1 (closed bars) inhibited SV40 early promoter and stimulated Mpl promoter fragments compared to chloramphenicol acetyl transferase (CAT; open bars). (C) Same as A for the Nptx1 locus. The presence of two G4C4 sites in the Nptx1 gene is indicated. (D) Same as B for the Nptx1 locus. Data are mean ± S.D. of four independent experiments. ***P < 0.001; **P < 0.01; *P < 0.05.