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. Author manuscript; available in PMC: 2015 May 13.
Published in final edited form as: Nat Commun. 2015 Apr 2;6:6411. doi: 10.1038/ncomms7411

Figure 7. Sox2 negatively regulates WWC1 by binding to its 3′UTR.

Figure 7

(a) Sox2 binds to the WWC1 3′UTR. Sox2 ChIP-Seq shows one peak (green) of Sox2-bound genomic sequence in the WWC1 3′UTR in SOX2-overexpressing osteoprogenitors29. Inset shows validation of ChIP-Seq in osteosarcoma cells by PCR analysis using primers flanking the Sox2-bidning site in the WWC1 3′UTR. (b) Analysis of WWC1 expression in mOS-482 cells expressing scrambled or two independent Sox2 shRNAs- A and B. Left panel: WWC1 mRNA expression normalized to actin levels. Middle panel: western analysis of WWC1. Right panel: luciferase assay. A 800-bp fragment of the WWC1 3′UTR containing Sox2-binding sites was cloned downstream of the luciferase-coding region in the pTK vector. Activity of the WWC1 3′ UTR luciferase construct was normalized to Renilla luciferase activity. *P<0.05 by t-test. Error bars represent mean ± s.d. (c) Analysis of WWC1 expression in primary osteopblasts cells expressing control or Sox2 lentivirus. Left panel: western analysis of WWC1. Middle panel: WWC1 mRNA expression normalized to actin levels. Right panel: luciferase assay. Activity of the WWC1 3′UTR luciferase construct was normalized to Renila luciferase activity. *P<0.05 by t-test. Error bars represent mean ± s.d.