Extended Data Fig. 5: Analyses of ELF4 movement from shoots to roots.

a, ELF4 and b, GFP proteins purified from bacteria and c, injected in shoots of elf4-1 mutant plants to examine rhythmic recovery in roots. d, e, Representative images showing fluorescence signals in roots of ELF4-ox-GFP scion and elf4-1 rootstock. Scale bars: 100µm. f, Gene expression analyses of ELF4 mRNA expression in WT and different ELF4-x3GFPs lines. Data are represented as the median ± max and min; 25-75 percentile. g, Hypocotyl length of different lines expressing ELF4-x3GFPs (E43GFP) (E43GFP1 n=34; E43GFP3 n=35) transformed into elf4-1 mutant plants. Hypocotyl length was also assayed for WT (n=27), elf4-1 (n=48) and plants over-expressing ELF4 fused to 1 GFP (E41GFP) (n=19). *** p-value<0.0001; two-tailed t-tests with 95% of confidence. Data are represented as the median ± max and min; 25-75 percentile. Circadian time course analyses of h, ELF4, and i, PRR9 mRNA expression by RT-QPCR in shoots of WT and ELF4-x3GFPs. j, Luminescence of elf4-1 scion into elf4-1 rootstocks elf4-1 (Sh)/elf4-1(Rt) (n=4) and its comparison with luminescence in WT (Sh)/WT(Rt) roots (n=5). k, Luminescence signals of elf4-1 (Sh)/elf4-1(Rt) from j, shown in a separate graph. Water instead of luciferin was added to the wells containing WT and elf4-1 shoots. h-k, Data are represented as the means + SEM. a-k, At least two biological replicates were performed for all experiments, with measurements taken from distinct samples grown and processed at different times.

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