Figure 1. FRS7 and FRS12 are circadian and photoperiodic-regulated genes.

(a) Circadian bioluminescence expression analysis of FRS7 and FRS12. Four-week-old Arabidopsis Col-0 wt protoplast cells from plants growing at 12L:12D cycle were transfected with ProFRS7:fLUC or ProFRS12:fLUC reporter constructs and transferred to constant red light at ZT9 for image acquisition with 2-h intervals for 1 week. Data represent the mean±s.e.m. (n=6, corresponding to six wells of protoplasts that were imaged and averaged for each time point of the figure). Two trials were performed, the results of which are shown in this panel and in Supplementary Fig. 2, respectively. White and grey regions indicate subjective light and dark period, respectively. (b) Diurnal oscillations of FRS7 and FRS12 transcript levels in Col-0 wt seedlings grown in SD or LD light conditions. ‘1’ represents the highest level of expression for a particular gene. Light and dark grey rectangles represent the dark period in SD and LD, respectively. Values represent the average expression of three biological replicates±s.e.m. (c,d) Immunoblot analysis showing the diurnal oscillation patterns of HA-tagged FRS7 and FRS12 proteins expressed through their respective native promoters in SD (c) and LD (d) growth conditions. White and black regions indicate light and dark period, respectively. (e) Comparison between SD and LD accumulation levels of HA-tagged FRS7 and FRS12 proteins expressed under control of their respective native promoters at ZT8 and ZT20. Uncropped versions of all immunoblot images presented in c–e are shown in Supplementary Fig. 15.