And fragments (1-6). Data shown as imply s.e.m. (n = three; panels k-n). Distinctive letters denote substantial differences (P 0.05, Duncan’s various range test). o, OsGRF4 activates pOsAMT1.2, pOsGS2, pOsNADH-GOGAT2 and pOsFd-GOGAT promoter::Luciferase fusion constructs in transient transactivation assays. Information shown as mean s.e.m. (n = three). P 0.05 as in comparison to manage group by two-sided Student’s t-tests.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsNature. Author manuscript; available in PMC 2019 February 15.Li et al.Pagea, Relative abundance of NO3- uptake transporter-encoding OsNRT1.1B, OsNRT2.3a and OsNPF2.four mRNAs. Abundance shown relative to that in NJ6 (=1). Information shown as imply s.e.m. (n = 3). Unique letters denote significant variations (P 0.05, Duncan’s multiple 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 = three). Distinctive letters denote significant differences (P 0.05, Duncan’s several variety 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. Data shown as mean s.e.m. (n = three). Different letters denote substantial variations (P 0.05, Duncan’s several variety test). i, OsGRF4 activates pOsNRT1.1B, pOsNRT2.3a, pOsNPF2.four, pOsNIA1, pOsNIA3 and pOsNiR1 promoter::Luciferase fusion constructs in transient transactivation assays. Information shown as imply s.e.m. (n = three) in all panels. A two-sided Student’s t-test was utilised to create the P values.Extended Information Figure 3. OsGRF4 regulates expression of many NO3- metabolism genes.Europe PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Information Figure 4. GA promotes GS and NR activities.a, GS activities in roots of 2-week-old rice plants treated with one hundred M GA (GA3) andor two M paclobutrazol (PAC), genotypes as indicated. Information shown as mean s.e.m. (n = three). A two-sided Student’s t-test was employed to produce the P values. b, GS activities in shoots of plants treated with GA andor PAC, genotypes and treatments as indicated within a. Data shown as imply s.e.m. (n = three). A two-sided Student’s t-test was used to generate the P values. c, NR activities in shoots of plants treated with GA andor PAC, genotypes and therapies as indicated within a. Information shown as imply s.e.m. (n = 3). A two-sided Student’s t-test was employed to produce the P values.Nature. Author manuscript; accessible in PMC 2019 February 15.Li et al.PageEurope PMC Funders Author Manuscripts Europe PMC Funders Author ManuscriptsExtended Information Figure 5. BiFC visualisation of SLR1-OsGIF1-OsGRF4 interactions.a, Information of constructs expressing OsGRF4 and Agents that act Inhibitors Related Products variants deleted for precise domains. OsGRF4 contains the QLQ (Gln, Leu, Gln) and WRC (Trp, Arg, Cys) Solriamfetol medchemexpress domains, positions as indicated. b, BiFC assays. Constructs expressing OsGRF4 or deletion variants (shown as within a) tagged together with the N-terminus of YFP have been co-transformed into tobacco leaf epidermal cells, together with constructs expressing OsGIF1 or SLR1 tagged with the C-terminus of YFP, respectively. Scale bar, 60 m. c, BiFC assays. Constructs expressing OsGRF1 or associated OsGRFs and OsGIFs family protein tagged using the N-terminus of YFP-tagged wereNature. Author ma.
