And fragments (1-6). Data shown as mean s.e.m. (n = three; panels k-n). Distinct letters denote important differences (P 0.05, Duncan’s numerous range test). o, OsGRF4 activates pOsAMT1.2, pOsGS2, pOsNADH-GOGAT2 and pOsFd-GOGAT promoter::Luciferase fusion constructs in transient transactivation assays. Information shown as imply s.e.m. (n = three). P 0.05 as in comparison with handle group by two-sided Student’s t-tests.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsNature. Author manuscript; readily available in PMC 2019 February 15.Li et al.Pagea, Relative abundance of NO3- uptake transporter-encoding OsNRT1.1B, OsNRT2.3a and OsNPF2.4 mRNAs. Abundance shown relative to that in NJ6 (=1). Data shown as mean s.e.m. (n = 3). Diverse letters denote important variations (P 0.05, Duncan’s several range test). b, Relative abundances of OsNIA1, OsNIA3 and OsNiR1 mRNAs encoding NO3- assimilation enzymes. Abundance shown relative to that in NJ6 (=1). Data shown as mean s.e.m. (n = 3). Different letters denote substantial differences (P 0.05, Duncan’s various range test). c-h, Flag-OsGRF4 mediated ChIP-PCR enrichment (relative to input) of GCGG-containing fragments (marked with ) from NO3- uptake transporter-encoding (c) OsNRT1.1B, (d) OsNRT2.3a and (e) OsNPF2.four gene promoters; NO3- assimilation enzymeencoding (f) OsNIA1, (g) OsNIA3 and (h) OsNiR1 gene promoters. Information shown as mean s.e.m. (n = 3). Diverse letters denote significant variations (P 0.05, Duncan’s numerous variety test). i, OsGRF4 activates pOsNRT1.1B, pOsNRT2.3a, pOsNPF2.four, pOsNIA1, pOsNIA3 and pOsNiR1 promoter::Luciferase fusion constructs in transient transactivation assays. Data shown as imply s.e.m. (n = 3) in all panels. A two-sided Student’s t-test was used to produce the P values.Extended Data Figure three. OsGRF4 regulates expression of numerous NO3- metabolism genes.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Data Figure four. GA promotes GS and NR activities.a, GS Af9 Inhibitors Related Products activities in roots of Eniluracil Autophagy 2-week-old rice plants treated with 100 M GA (GA3) andor two M paclobutrazol (PAC), genotypes as indicated. Data shown as mean s.e.m. (n = 3). A two-sided Student’s t-test was employed to create the P values. b, GS activities in shoots of plants treated with GA andor PAC, genotypes and treatments as indicated in a. Information shown as imply s.e.m. (n = 3). A two-sided Student’s t-test was employed to create the P values. c, NR activities in shoots of plants treated with GA andor PAC, genotypes and therapies as indicated within a. Data shown as mean s.e.m. (n = 3). A two-sided Student’s t-test was utilized to generate the P values.Nature. Author manuscript; available in PMC 2019 February 15.Li et al.PageEurope PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Data Figure 5. BiFC visualisation of SLR1-OsGIF1-OsGRF4 interactions.a, Specifics of constructs expressing OsGRF4 and variants deleted for specific domains. OsGRF4 contains the QLQ (Gln, Leu, Gln) and WRC (Trp, Arg, Cys) domains, positions as indicated. b, BiFC assays. Constructs expressing OsGRF4 or deletion variants (shown as inside a) tagged with the N-terminus of YFP were co-transformed into tobacco leaf epidermal cells, together with constructs expressing OsGIF1 or SLR1 tagged with all the C-terminus of YFP, respectively. Scale bar, 60 m. c, BiFC assays. Constructs expressing OsGRF1 or related OsGRFs and OsGIFs loved ones protein tagged with all the N-terminus of YFP-tagged wereNature. Author ma.