Fig 1.
Design of luciferase reporter T87 cells.
(A) A schematic diagram showing the vector constructs used in this study. 35S, sec, and T indicate the 35S promoter, secretory signal, and terminator polyadenylation signal, respectively. (B) Box plot of luminescence intensities for luciferase reporter Arabidopsis thaliana T87 cells on addition of furimazine or d-luciferin. Nano-Glo luciferase assay system and Dual-Glo luciferase assay system were used for NLuc and FLuc, respectively. The data are from three individual experiments with biological octuplicates; n = 24. Values indicate averages.
Fig 2.
Effect of pH conditions on GeNL reporter activity.
The luminescence intensities of GeNL and secGeNL reporter cells were determined on addition of furimazine under different pH conditions (5.0, 5.8, 6.6, 7.4, and 8.2). Values shown are the mean ± SE; n = 3.
Fig 3.
Detection of reporter cell luminescence through a layer of T87 cells.
(A) Normalized emission spectra of NLuc (blue) and GeNL (green) reporter Arabidopsis thaliana T87 cells on addition of furimazine. The emission spectra of NLuc (blue dotted line) and GeNL (green dotted line) cells on the addition of coelenterazine h are also shown. The gray shade indicates the absorbance spectrum of wild-type T87 cells. The spectra were normalized to peak height for each reporter cell type in the range 400 to 700 nm. (B) Luminescence images of NLuc (blue) and GeNL (green) reporter T87 cells on addition of NLuc luciferin through a layer of wild-type T87 cells. (C) Transmittance rates of reporter cells covered with a layer of T87 cells. Transmittance rates were calculated as the transmitted luminescence intensity in the presence of a layer of wild-type T87 cells, divided by that in the absence of a layer. The values shown are the mean ± SE; n = 4. Student’s t test was performed. ***p < 0.001.
Fig 4.
Luminescence imaging of GeNL expressing Arabidopsis thaliana.
(A) Fluorescence image of GeNL-expressing Arabidopsis thaliana in a 10 cm dish. The image was obtained using a GelGreen filter (560/50 nm). (B) Luminescence image of GeNL-expressing A. thaliana in a 48-well plate on addition of furimazine. WT indicates wild-type A. thaliana plants.