(A) Heatmap of the commonly deregulated genes upon silencing of KLF5 in SW480. Sixty-four genes were significantly deregulated in the KLF5-specific RNAi signature. Note that two different siRNAs against this gene result in a very reproducible deregulation of the same signature genes. Among the most downregulated (blue) and upregulated (red) genes are the transcription factors FGF18, RUNX1, and SOX2, as well as the WNT-pathway member WNT10A. (B) Average expression levels of KLF5, FGF18, WNT10A, and SOX2 in cell lines, primary tumors and in SW480 before and after silencing of KLF5. Color code on the right.
(C) Western blot analysis of expression levels of KLF5 and SOX2 after silencing of KLF5 with two different siRNAs. Controls: untransfected cells (WT) and cells transfected with AllStarNeg. The profound downregulation of KLF5 and upregulation of SOX2 are consistent with the gene expression analysis.