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. 2019 Feb 7;9:1629. doi: 10.1038/s41598-018-38310-y

Figure 2.

Figure 2

Aberrant Wnt/β-catenin signaling activates MSX1 expression in human cells. (A) Quantitative RT-PCR analysis of the MSX1 mRNA expression level in HEK293 cells upon treatment with Wnt3a-conditioned medium (left diagram) or with GSK3 inhibitor BIO (right diagram). The diagrams show expression levels of the indicated gene in Wnt3a- or BIO-treated cells relative to the levels determined in control cells without treatment. (B) MSX1 is upregulated in STF cells expressing truncated APC. Quantitative RT-PCR analysis of STF cells producing wild-type or truncated APC proteins. The mutant APC gene was generated by targeting exon 10 or exon 15 using TALENs or the CRISPR/Cas9 system, respectively. The diagram shows corresponding Ct values upon normalization to β-actin mRNA levels. (C) Decreased expression of MSX1 upon siRNA-mediated knock-down of β-catenin mRNA. Quantitative RT-PCR analysis of SW480 and SW620 CRC cells and STF cells expressing a truncated APC variant generated by exon 10 targeting transfected with β-catenin-specific siRNA. The gene expression level in cells transfected with non-silencing siRNA was set to 1. Each analysis depicted in panels A, B, and C was performed at least twice; results of one representative experiment for the given experimental setup are shown. PCR reactions were run in triplicates; error bars indicate SDs. The amounts of total RNA in individual samples were normalized to β-actin expression, the results for additional housekeeping gene ubiquitin B (UBB) are shown. Corresponding Ct values are given in Supplementary Table S3.