Proteome Profiler Human Pluripotent Stem Cell Array Kit …

siRNA-mediated Knockdown of PDGF R alpha, PDGF R beta, and cAbl Upregulates Expression of Pluripotent, Mesoderm, and Endoderm Markers. Small interfering RNAs (siRNAs) were transfected into mesenchymal stem cells by electroporation. Knockdown was verified by RT-PCR and Western blot analysis (data not shown here). Cell lysates from siRNA-treated cells were used in the Proteome Profiler Human Pluripotent Stem Cell Antibody Array (Catalog#ARY010) to examine relative expression levels of mesoderm, endoderm, and pluripotent markers. The images represent nitrocellulose membranes following chemiluminescent detection of bound analytes. Each of the array markers has duplicate spots, which are boxed to highlight their identity. A. Scrambled (Scr) siRNA-treated mesenchymal stem cells. B. Knockdown of PDGF R alpha upregulated pluripotent markers (Oct-3/4, Nanog, SOX2, and E-Cadherin) as well as markers of the mesoderm and endoderm compared to Scr. C. Knockdown of PDGF R beta upregulated expression of Snail, SOX17, VEGF R2, Oct-3/4, and Nanog compared to both Scr and PDGF R alpha knockdowns. D. Knockdown of cAbl upregulated pluripotent markers (Nanog, Oct-3/4, E-Cadherin, and SOX2) compared to Scr in addition to markers of the endoderm and mesoderm.

Adapted from Ball, S.G. et al. (2012) Stem Cells 30:548.

Abbreviations: AFP, alpha-Fetoprotein; GAPDH, Glyceraldehyde 3-Phosphate Dehydrogenase; GSC, Goosecoid; HCG, Human Chorionic Gonadotropin; HNF, Hepatocyte Nuclear Factor; PDGF R, Platelet-derived Growth Factor Receptor; Scr, scrambled; VEGF R, Vascular Endothelial Growth Factor Receptor.

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Proteome Profiler Human Pluripotent Stem Cell Array Kit ...