Figure 4
Regulation of cAMP increase with red light.
(A) Red light-inducible dimerization of the PhyB/PIF6 system allows Gαs to translocate to the plasma membrane and trigger cAMP release owning to the activation of adenylyl cyclase (AC). (B) Constructs for PhyB-FusionRed-CAAX and PIF6-Gαs. (C) Time-lapse luminescence intensity of samples under red illumination at 660 nm (red) or far-red illumination at 735 nm (far-red), with (PCB (+)) or without (PCB (−)) addition of PCB. The increment of luminescence intensity was only observed in the samples under red illumination in the presence of PCB. The upper table means the normalized intensity ratio changes between red PCB (+) and far-red PCB (+). (D) Relationship between PCB concentration and the luminescence intensity. (E) Power dependency of the luminescence intensity under illumination at 660 nm. (F) Result of the bioluminescence assay after optimization of several parameters including PCB concentration (50 μM), illumination time (14 hours) and light power (red: 0.15 mW/cm2; far-red: 1.5 mW/cm2). The inset is a partially enlarged view of mock, which means a negative control in the absence of PhyB-FusionRed-CAAX. The error bars indicate standard deviation from three individual samples (n = 3).
