Fig. 1: BLI of NS/PC-derived cells stimulated with high potassium in vitro.
From: A non-invasive system to monitor in vivo neural graft activity after spinal cord injury
a Schematic illustration of the E-SARE-Venus-AkaLuc (ESAL) construct, which was used to express the Venus-fused AkaLuc luminescent enzyme under the control of the promoter E-SARE. When a neuron was activated, the promoter E-SARE drove high expression of the downstream reporter gene Venus-AkaLuc. b Comparative bioluminescence imaging (BLI) of in vitro-cultured cells stimulated with 50 mM KCl for 6 h (on the right, n = 4) or without stimulation (on the left, n = 4) on the left. We prepared n = 8 wells from two independent tertiary neurospheres from the same embryoid body (EB). The colour of the bars indicates the total bioluminescence radiance (photons/sec/cm2/str). The steradian (str) is the unit of the solid angle. c Quantitative analyses of the relative BLI signal intensity of ESAL-NS/PC-derived cells with or without the addition of 50 mM KCl in vitro (n = 4 each). The values are the mean ± standard error of the mean (SEM): **p < 0.01. A two-sided unpaired Student’s t test was performed. T value and degrees of freedom: t (6) = 11.59, p = 2.5 × 10−5. d Microscopic bright-field image and Venus fluorescence image of ESAL-NS/PC-derived cells with or without the addition of 50 mM KCl in vitro. Scale bar, 50 μm. e The results of qPCR analyses of the gene expression of Venus, ARC, and FOS in cells within the same well as shown above (n = 4 each). The values are the mean ± SEM: *p < 0.05, **p < 0.01. A two-sided unpaired Student’s t test was performed. Individual t values and degrees of freedom: Venus t(6) = 3.034, p = 0.023. Arc t(6) = 2.705, p = 0.035. Fos t(6) = 7.057, p = 4.1 × 10−4. f, g Representative images of Venus-expressing differentiated cells from NS/PCs stained for panembryonic lethal abnormal vision-like (pan-ELAVL) (neurons) with or without the addition of 50 mM KCl (f); stained for human glial fibrillary acidic protein (GFAP) (astrocytes), 2ʹ, 3ʹ -cyclic nucleotide 3ʹ-phosphodiesterase (CNPase) (oligodendrocytes), Nestin and Ki-67 (immature cells) with the addition of 50 mM KCl (g). Scale bar, 20 μm. h Percentage of cells positive for cell type-specific markers among the Venus+ cells after stimulation with 50 mM KCl.
