For 'KHM221' plants, 50 μmol m−2 s−1 white, blue, and red light conditions, and darkness were applied with duplicates.
During the dark period, blood sampling took place under red light conditions.
Dark control animals (n = 3) received L-AP4 in one eye and saline in the other eye under similar dim red light conditions, but after recovering from anesthesia the animals were returned to the dark and killed 3 h later.
All behavioural tests were conducted during the first 2 hours of the dark phase (corresponding to the consumption measurement interval) either at reduced illumination (mHb: 30 lx) or under red light conditions (all other tests).
Under isofluorane anesthesia, animals received a unilateral intraocular injection of L-AP4 (1 µl; 100 µM final concentration) under dim red light (approximately 5 lux); the contralateral eye served as a control and was injected with 1 µl of vehicle (0.9% sterile saline) under the same dim red light conditions.
QTL for hypocotyl length in this region, on the bottom of chromosome 4, have been previously identified in another RIL population in white, blue and red light conditions, suggesting that this region of the genome may harbor a QTL important for seedling light responsiveness [38].
In addition, the enhanced FAA phenotype of KO mice persists during two days of total food deprivation and also in constant dim red light conditions, indicating that FAA in these mice is not driven by an hourglass mechanism or attributable to altered sensitivity to light.
Since hyperlocomotion still appeared under red light conditions in the open field test in BL/6N mice (data not shown), our CMS protocol seems to exert additional effects on reactivity to a test situation that lie beyond an increased sensitivity to even moderate illumination conditions.
We combined the pif-mutant data of Khanna et al. [28], Al-Sady and colleagues [46] as well as Leivar et al. [29] with our over-expressing lines phyB-GFP-1 to 4 relative to the phyB level of Col WT, since all experiments were conducted under saturating red light conditions for four days.
