FIGURE 3.

GmERD15 localizes to the nucleus of tobacco leaf cells and binds to the NRP-B promoter in vivo. A, confocal fluorescence image of transiently expressed GmERD15-YFP in epidermal cells of tobacco leaves. Tobacco leaves were infiltrated with Agrobacterium tumefaciens carrying ³⁵S::GFP-NSP or ³⁵S::GmERD15-YFP DNA constructs and observed by confocal microscopy 3 days after agroinoculation. The white arrows show the nuclear positions of the nuclear viral protein NSPs (left panel) and GmERD15 (right panel). The black arrows in the bottom right panel show the co-localization signal of GmERD15 with the nuclear marker NSP. B, immunoblotting of nuclear fractions of tobacco leaves expressing YFP-GmERD15. Soluble (SF) and nuclear fractions (NF) of tobacco leaves expressing YFP-GmERD15 were immunoblotted with anti-GmERD15 antibodies. The open arrow indicates the position of purified His-GmERD15, and the black arrow indicates the position of YFP-GmERD15. M is molecular mass. C, ChIP with tunicamycin-treated soybean suspension cells using anti-GmERD15 antibodies. The 187-bp DNA fragment of NRP-B 5′-flanking sequences was detected by PCR amplification when immunoprecipitation was mediated by anti-GmERD15 antibodies (lane 2) but not by unrelated anti-polymerase (Pol) II antibodies (lane 4) or IgG (lane 3). Lane 1 represents the amplification from input total DNA, and M represents the molecular mass standards.