Journal: Neural Plasticity
Article Title: Sphingosine 1-Phosphate Receptor 1 Modulates CNTF-Induced Axonal Growth and Neuroprotection in the Mouse Visual System
Figure Lengend Snippet: Modulation of S1PR1 expression by the CNTF/Stat3 pathway in optic nerve-injured retinae. (a, b) S1PR1 expression changes were monitored 3 d after ONC in retinae infected with ShH10.CNTF or ShH10.Empty, a control vector that was deprived of cDNA sequence. ShH10 viruses preferentially infected Müller glia in the retina [ 16 ]. P-Stat3 and S1PR1 were markedly increased by ShH10.CNTF in RGC somata identified using β 3-tubulin as a specific marker. (c) Five days after ONC, qRT-PCR measurements showed that the infection of RGCs with AAV2.Stat3 significantly increased the mRNA level of S1pr1 compared with control AAV2.GFP vector. ShH10 and AAV2 viruses were intravitreally injected 4 weeks before ONC. Three mice were analyzed/grouped. Statistics: one-way ANOVA; ∗ p
Article Snippet: Primary antibodies were rabbit anti-β 3-tubulin (1 : 1000; ab18207, Abcam), mouse anti-β 3-tubulin (1 : 1000; G712A, Promega, Madison, WI, USA), rabbit anti-S1PR1 (1 : 50; ASR-011; Alomone Labs, Jerusalem, Israel), rabbit anti-S1PR1 (1 : 200; PA1-1040, Life Technologies), and rabbit anti-phospho-Stat3 (1 : 100; 9131, Cell Signaling, Whitby, ON, Canada).
Techniques: Expressing, Infection, Plasmid Preparation, Sequencing, Marker, Quantitative RT-PCR, Injection, Mouse Assay