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. 2008 Oct 6;183(1):37–48. doi: 10.1083/jcb.200804030

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© 2008 Wang et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 5. — Notch signaling is active but cannot induce terminal differentiation in the absence of AP-2α and AP-2γ. (A) Real-time PCR analyses of WT and DKO 1°MK transduced with either an IRES-GFP empty vector (control) or an IRES-GFP retroviral vector encoding the NICD. After 36 h, transduced cells were FACS enriched and K14, K1, and K10 mRNA levels were measured. Fold change upon NICD induction was compared with the control vector. (B) Same experiment as in A, but documenting the mRNA levels of Hes1, Hey1, and RBP/J. (C) Same experiment as in A, but showing that AP-2 mRNA levels are only modestly affected by activated Notch. (D) RBP/J^lox/lox 1°MK were cultured in duplicate in low Ca²⁺ medium, and then transduced with either an adenovirus expressing a cytomegalovirus-Cre recombinase (Cre) or cytomegalovirus-GFP (control). At t = 2 d, one set of transduced cells were switched to high Ca²⁺ to induce differentiation. FACS was performed at t = 4 d and mRNAs were subjected to real-time PCR analyses with the primers indicated. Error bars represent standard deviation for three or more independent experiments.