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. 2014 Jul 17;5(7):e1332. doi: 10.1038/cddis.2014.301

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Copyright © 2014 Macmillan Publishers Limited

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 3.0 Unported License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/3.0/

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Fluvoxamine (Flv) induces the expression of sigma-1 receptor (Sig-1R). (a) Flv (10 μg/ml) was added to Neuro2a cells, and Sig-1R protein and GAPDH protein levels were monitored over the time course (h) shown in the figure. (b) The amount of Sig-1R and GAPDH protein in (a) was quantified with densitometry. Relative intensity of Sig-1R/GAPDH increased significantly in response to Flv treatment. The 0 h time point was normalized to 1. Values shown are the mean±S.D. (*P<0.05, Student's t-test; n=3). (c) Sig-1R and β-actin mRNA expression in Neuro2a cells treated with 10 μg/ml Flv was measured by semiquantitative RT-PCR over the time course shown in the figure. The Sig-1R mRNA level increased by ∼3.8-fold after Flv treatment for 24 h. (d) Sig-1R and β-actin mRNA expression in Neuro2a cells treated with 10 μg/ml Flv was determined by quantitative RT-PCR over the time course shown in the figure. The Sig-1R mRNA level increased by 32% and 28% after Flv treatment for 12 h and 24 h, respectively. The 0 h time point was normalized to 1. Values are the mean±S.D. (*P<0.05, Student's t-test; n=3). (e) The reporter assay system shown in the figure was constructed. The Sig-1R promoter region (-582 to -156) was fused to a firefly luciferase plasmid (pGL4.12[luc2CP]) and relative luciferase activities were measured. Luciferase activity increased significantly over the time course shown in response to treatment with 10 μg/ml Flv. Values are the mean±S.D. (*P<0.05, Student's t-test; n=3)