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. 2015 Aug 15;29(16):1677–1682. doi: 10.1101/gad.261677.115

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© 2015 Chen et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 5. — Neurofibromin/Jagged1 regulation of astrocyte differentiation requires MEK-mediated Smad3 expression. Increased Jagged1 transcription (A) and protein levels (B) in Nf1^−/− NSCs were reduced to wild-type levels following 5 nM PD901 treatment. PD901 treatment of Nf1^−/− NSCs had no effect on β-catenin activity or YAP expression but reduced Smad3 expression by Western blotting (B) and immunocytochemistry (C). (D) Smad3 knockdown restored Jagged1, NICD, and Hes1, but not Hes5, expression to wild-type levels. (E) The astrocyte, oligodendrocyte, and neuronal differentiation defects observed in Nf1^−/− TVZ NSCs were restored to near wild-type levels following Smad3 knockdown. (F) MEK inhibition (5 nM PD901) reduced Olig2, but not Olig1, expression by qRT–PCR and Western blotting. Smad3 knockdown reduced Olig2 expression by Western blotting (G) as well as the percentage of Olig2⁺ cells within the Nf1^−/− neurospheres (H). (I) Proposed model of neurofibromin/RAS regulation of NSC growth and multilineage differentiation. (veh) Vehicle. Error bars denote mean ± SD. Bar, 100 μm. (*) P < 0.05; (**) P < 0.01.