Fig. 3: Subfunctionalisation of soybean E1 proteins. | Nature Communications

Fig. 3: Subfunctionalisation of soybean E1 proteins.

From: Subfunctionalisation and self-repression of duplicated E1 homologues finetunes soybean flowering and adaptation

Fig. 3

a Alignment of full-length E1 orthologues from Wm82-E1. NLS represents nuclear-localisation signal. The numbers indicate every 20 residues. ‘Ω’ depicts the N-terminus peptide of E1 family proteins. b Schematic of the constructs used to assay subcellular localisation of soybean E1 proteins by confocal microscopy in transiently transgenic N. benthamiana leaves. GFP (Green Fluorescent Protein) was fused to C-termini and constructs were under the control of the CaMV35S (Cauliflower Mosaic Virus 35S) promoter. ‘Ω’ depicts the N-terminus peptide of E1 family proteins. c Confocal-microscopy analysis of subcellular localisation of soybean E1 proteins and their variants as summarised in (b). Scale bar = 25 μm. Three independent biological replicates were performed. d ddPCR comparison of E1, E1La and E1Lb expression in Wm82-E1 at ZT 4 under long-day conditions at 20 DAE in a growth chamber. Data are the mean ± S.D. of n = 3 biological replicates. The two-sided Student’s t-test was used to generate the P values. e ddPCR comparison of E1La and E1Lb expression in Wm82-E1 and Wm82 (e1-as) backgrounds at ZT 4 under long-day conditions at 20 DAE in a growth chamber. Data are the mean ± S.D. of n = 3 biological replicates. The two-sided Student’s t-test was used to generate the P values. f ddPCR comparison of E1 expression in Wm82-E1 and Wm82 (e1-as) backgrounds at ZT 4 under long-day conditions at 20 DAE in a growth chamber. Data are the mean ± S.D. of n = 3 biological replicates. The two-sided Student’s t-test was used to generate the P values. ChIP–qPCR assay for enrichment of E1–3xFLAG at the promoter of E1 (g), E1La (h) and E1Lb (i). ELF1b served as a negative control. Schematics of promoters and coding regions are shown on the left with the amplified regions depicted (P1–3). ‘N’ depicts negative control. Data are the mean ± S.D. of n = 4 biological replicates. The two-sided Student’s t-test was used to generate the P values. ns indicate no significant difference (P > 0.05). Source data are provided as a Source Data file.

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