Journal: International Journal of Molecular Sciences
Article Title: Transcriptional Regulation of the Synaptic Vesicle Protein Synaptogyrin-3 ( SYNGR3 ) Gene: The Effects of NURR1 on Its Expression
doi: 10.3390/ijms23073646
Figure Lengend Snippet: Effects of Nurr1 activator C-DIM12 on protein expression in SH-SY5Y cells and the effect of activator activity assayed in real time. ( A ) (i) total cell lysates from vehicle-treated and C-DIM12 treated SH-SY5Y cells were detected by anti-SYNGR3 and anti-Actin antibodies. The Western blots showed bands at 25 kDa corresponding to SYNGR3 and bands at 43 kDa corresponding to actin. SYNGR3 protein expression increased in SH-SY5Y cells treated with C-DIM12 compared with that in cells treated with vehicle. (ii) quantitative measurement of the SYNGR3 expression levels in C-DIM12 and vehicle-treated cells. ( B ) (i) schematic diagram of the luciferase reporter plasmids cloned with the 2029-bp 5′-flanking region of pGL3-hSYNGR3–1870/ATG containing three NBRE-like elements. SH-SY5Y cells were transiently transfected with the pGL3-hSYNGR3–1870/ATG construct containing three NBRE-like elements. Real-time luciferase activity was measured from 12 h to 36 h following treatment with vehicle and C-DIM12 (10μM) in SH-SY5Y cells. (ii) Cumulative luciferase activity was calculated from the area under the curve (AUC) of the real-time luciferase activity of pGL3-hSYNGR3–1870/ATG in SH-SY5Y cells treated with vehicle and C-DIM12. Values are mean ± SEM of six independent experiments. *** indicates significance at p < 0.01 as compared to vehicle-treated controls. * indicates significance at p < 0.05 compared with the level in vehicle-treated controls. The bars indicate mean ± SEM of the three independent experiments.
Article Snippet: SH-SY5Y cells were treated with a NURR1 activator, C-DIM12 (Bio-Techne Hong Kong Ltd., Hong Kong) to induce NURR1 expression/activation.
Techniques: Expressing, Activity Assay, Western Blot, Luciferase, Clone Assay, Transfection, Construct