Molecular Neurobiology
Supplementary Materials
Contra-directional coupling of Nur77 and Nurr1 in Neurodegeneration: A novel mechanism for Memantine-induced anti-inflammation and anti-mitochondrial impairment
Xiaobo Wei MD 1†, Huimin Gao MD 1†, Jing Zou MD 1†, Xu Liu MD 1, Dan Chen MD1, Jinchi Liao MD 1, Yunqi Xu MD 1, Long Ma Ph.D.2, Beisha Tang MD, Ph.D.2, Zhuohua Zhang Ph.D.2, Xiang Cai Ph.D 3, Kunling Jin MD, Ph.D 4, Ying Xia MD, Ph.D 5*, Qing Wang MD, Ph.D 1*
1 Department of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University, Tianhe Road 600, Guangzhou, Guangdong 510630, P.R. China
2 The State Key Laboratory of Medical Genetics, Central South University,Changsha, Hunan 410078, China
3 Institute of Neuroscience and the Second Affiliated Hospital of Guangzhou Medical University, 250# Changgang East Road, Guangzhou, 510260, China
4 Department of Pharmacology and Neuroscience, Institute for Aging and Alzheimer's Disease Research, University of North Texas Health Science Center, Fort Worth, TX 76107, USA
5 Department of Neurosurgery, The University of Texas Medical School at Houston, Houston, TX 77030, USA
Correspondence and reprint request to:
Prof. (Dr.) Qing Wang MD, Ph.D
Vice-Head, Department of Neurology,
Director of Neurological Research Lab,
The Third Affiliated Hospital of Sun Yat-Sen University,
Tianhe Road 600#, Guangzhou, Guangdong 510630,
P.R. China
Phone: +86-20-85253275； FAX: +86-20-85253117
Email:
Supplementary Figure 1:
Fig. S1 Treatment with 6-OHDA induced time- and concentration- dependent damage to PC12 cells
PC12 cells were subjected to different concentrations of 6-OHDA (0 μM, 50 μM, 100 μM, 150 μM , 200 μM), for various time points (6 h, 12 h, 24 h, 48 h). The viability of the PC12 cells was analysed via the CCK8 assay. The results are expressed as a relative ratio of the blank group and are expressed as the mean ± SEM of 5 independent experiments. ***p<0.001, one-way ANOVA followed by Bonferroni’s comparison post hoc analysis (Bonferroni’s t-test) 24 hr 100 μM 6-OHDA-treated group vs. the blank group. When the cells were incubated with 100 μM 6-OHDA for 24 h, the relative cell viability decreased to 52.49%. This dose and time point were chosen for the subsequent experiments; thus, we only indicated the significant difference (***) for this group in the bar chart.
Supplementary Figure 2:
Fig. S2 Memantine mediates survival/neurogenesis in 6-OHDA-lesioned PC12 cells via the Nurr1/BDNF/PI3K/AKT pathway
(A-B) After exposure to 6-OHDA (100 μM) with/without memantine (10 μM) for 24 h, total cellular lysates of PC12 cells were extracted and analyzed by immunoblot for Nurr1, BDNF, PI3K, p-PI3K, AKT, p-AKT and β-actin protein levels. (C-D) The diagram shows the relative quantitation of the specific protein levels of (A-B) compared with that of β-actin, showing that 6-OHDA consistently downregulated the expressions of Nurr1, BDNF, PI3K, p-PI3K, AKT, and p-AKT, which was partly attenuated by the addition of memantine. The data are expressed as the relative ratios of the blank group, which were set to 1.0 and are expressed as the mean ± SEM of 5 independent experiments. ***p<0.001, Bonferroni’s t-test vs. the blank group; ###p<0.001, Bonferroni’s t-test vs. the 6-OHDA group.
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