Fig. 3: Molecular interaction between Hd6-Hd2 module and N.

a Profile of Hd6 transcripts in response to different N concentrations in shoots. NIL(Hd6) seedlings were treated with different concentrations of NH₄Cl as indicated after pretreatment of 0.4 mM NH₄Cl for 3 days. Shoot parts were harvested as indicated hours (hr) after the N treatment for RNA extractions. mRNA levels were normalized by the copy numbers of Actin1 transcript. The mean values of 0.1 mM sample were set as 1. qPCR was performed in triplicate, and values with the SD are shown. There was no significant difference (P > 0.05) based on Tukey’s HSD test. b Profile of Hd6 protein in response to different N concentrations for 24 h. Sample preparation was essentially same as that in (a). Representative image was shown. Relative amounts of Hd6 protein were normalized to the values of the loading control, β-tubulin (β-Tub) and set value of 0.1 mM sample as 1 in the bottom of image. c Profile of Hd2 transcripts in response to different N concentrations in shoots. Samples correspond to that in (a). d Accumulation of 3FLAG-Hd2 in response to different concentrations of N source. Immunoblotting of 3FLAG-Hd2 was performed with crude extracts from 35S:Hd6/Ubq:Ω:3FLAG-Hd2 and Ubq:Ω:3FLAG-Hd2 seedlings treated with NH₄Cl. Seedlings were treated with the indicated N concentrations for 24 h after pretreatment of a lower concentration for N starvation. Immunoblotting with anti-β-Tub antibody was used for the loading control. Each experiment was repeated independently for at least 3 times with similar results. e, f mRNA levels of Hd6 (e) and Hd2 (f) in transgenic plants. The mRNA levels were normalized by that of Ubq. Mean values with the SD were obtained by three independent qPCR reactions. Complementation of Hd6 and Hd2 function by introduction of functional Hd6 and Hd2 into NIL (hd6 hd16). Source data are provided as a Source Data file.