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. Author manuscript; available in PMC: 2010 Jun 4.
Published in final edited form as: Nat Cell Biol. 2009 Dec 13;12(1):31–9. doi: 10.1038/ncb2001

Figure 2.

Figure 2

TLX activates Wnt/β-catenin/cyclin D1. (a) TLX potentiates a β-catenin-responsive reporter gene. The β-catenin-responsive reporter gene Topflash was transfected into CV-1 cells along with β-catenin or TLX, or both, in the presence of a control siRNA (C) or Wnt7a siRNAs (siWnt7a-1 and siWnt7a-3). Reporter luciferase activity was measured and normalized with β-galactosidase activity. Error bars show s.d.; assays were performed in triplicate. (b) Knockdown of Wnt7a expression with Wnt7a-specific siRNAs as revealed by luciferase assays. Wnt7a-specific siRNAs 1, 2 and 3 (si1, si2 and si3) were co-transfected with a luciferase reporter gene upstream of Wnt7a cDNA. The knockdown effect was determined by relative luciferase activity. GFP siRNA was included as a control (C). Error bars show s.d.; assays were performed in triplicate. Asterisk, P < 0.005 by Student’s t-test. (c) Expression of Wnt7a and cyclin D1 in TLX siRNA (siTLX)-treated adult mouse neural stem cells as revealed by RT–PCR analysis. Actin was included as a loading control. (d) Expression of Wnt7a and cyclin D1 in TLX-expressing 3T3 (3T3-TLX) cells and parental 3T3 cells as revealed by northern blot analysis. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was included as a loading control. (e) Expression of TLX and cytosolic β-catenin in 3T3-TLX and parental 3T3 cells demonstrated by western blot analysis. Uncropped images of blots in (c)–(e) are shown in Supplementary Information, Fig. S5.