Transcriptomic Profiling of Schwann Cells with Sox2 Overexpression

Schwann cells were infected with control GFP or Sox2/GFP adenovirus (Le et al., 2005 (link), Parrinello et al., 2010 (link)) for 24 hours in defined medium (DM) and then changed to DM without virus. After 48 hours of adenovirus infection, total RNA was purified with miRNeasy mini kit (QIAGEN, 217004) and on-column DNase digestion was also performed. Triplicate mRNA samples purified from GFP or Sox2/GFP adenovirus-infected Schwann cells were analyzed on an Affymetrix 1.0 ST Rat Exon array by the Patterson Institute for Cancer Research (Manchester, UK). Affymetrix data was analyzed using the Affymetrix Power Tools software. Regulated transcripts were identified using a false discovery rate controlled p value of < 0.05. The putative functional role of Sox2 targets was then analyzed by Gene Ontology analysis with the Gene Map Annotator and Pathway Profiler (GenMAPP) software package. The 9 classic axon guidance receptors were identified from axon guidance signaling pathways.

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Dun X.P., Carr L., Woodley P.K., Barry R.W., Drake L.K., Mindos T., Roberts S.L., Lloyd A.C, & Parkinson D.B. (2019). Macrophage-Derived Slit3 Controls Cell Migration and Axon Pathfinding in the Peripheral Nerve Bridge. Cell Reports, 26(6), 1458-1472.e4.

Publication 2019

miRNeasy mini kit Power Tools software

Adenovirus Adenovirus infection Axon guidance pathways Cancer Digestion Dnase Exon Gene map Mrna Schwann cells Sox2 Virus

Corresponding Organization : University of Plymouth

Other organizations : University of Bath, MRC Laboratory for Molecular Cell Biology, University College London

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Variable analysis

independent variables

Infection with control GFP adenovirus
Infection with Sox2/GFP adenovirus

dependent variables

Transcripts regulated by Sox2 overexpression
Functional role of Sox2 targets analyzed by Gene Ontology

control variables

Defined medium (DM) used for cell culture
Duration of adenovirus infection (24 hours)

controls

Positive control: Infection with control GFP adenovirus
Negative control: Not explicitly mentioned

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