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. Author manuscript; available in PMC: 2012 May 17.
Published in final edited form as: Cancer Cell. 2011 May 17;19(5):601–612. doi: 10.1016/j.ccr.2011.03.010

Figure 2. Wnt autocrine activation in human sarcomas occurs commonly and is associated with several pathway aberrations.

Figure 2

(A) Effects of DKK1 on Wnt/β-catenin signaling in human sarcoma cells. Cells were treated with DKK1 for 3 hrs followed by analysis of uncomplexed β-catenin levels as described in Fig. 1A.

(B) Wnt activated sarcoma lines resistant to DKK1 inhibition were stably infected with human FRP followed by DKK1 treatment and immunoblot analysis as described in A.

(C) Real-time PCR analyses for various canonical Wnt ligands were conducted on total RNAs. Normalized values are represented relative to those in hMSC.

(D) Protein extracts from whole cell lysates of different human sarcoma lines and hMSCs were subjected to immunoblot analysis.

(E) Real-time PCR was performed on genomic DNAs extracted from the indicated sarcoma lines to analyze LRP5 gene copy number. SUV420H1 and GAL are two flanking genes 5′ and 3′ of LRP5, respectively. Albumin was used as a standard for diploid gene copy number. Normalized values are represented relative to those in normal human epithelial cells.

Error bars indicate SD of mean values obtained from triplicates and are representative of two independent experiments. See also Figure S2.