(A) Significantly upregulated Wnt genes comparing SW1463RES to SW1463. Differentially expressed genes (B) between SW1463, and SW1463 six hours after exposure to a single dose of 4 Gy of X-rays (SW1463-4 Gy). Differentially expressed genes (C) between SW1463RES, and SW1463RES six hours after exposure to a single dose of 4 Gy of X-rays (SW1463RES-4 Gy).

<p>Supplementary Figure S1 PDF file - 1274K, Schematic presentation of the workflow of experiments and analytical procedures</p>

Supplementary Figure S3 PDF file - 2243K, RNAi analysis and measurement of cell viability of amplified and overexpressed chromosome 13 candidate genes

Schematic model of the chemoradiation treatment after Wnt-3a stimulation.

<p>Supplementary Figure S9 PDF file - 1478K, Expression levels of the LNX2 module</p>

Supplementary Table S1 PDF file - 435K, ist of RT-PCR primers used for validation

Western blot analysis of (A) SW480, (B) SW837, and (C) RPE-1 cells after exposure to various doses of XAV-939 and DMSO as negative control.

Supplementary Table S4 PDF file - 358K, List of filtered genes used in our experimental setup

Supplementary Table S2 PDF file - 589K, List of siRNAs

Supplementary Figure S2 PDF file - 1800K, Corroboration of gene expression levels by qRT-PCR

Radiation sensitization effect of XAV-939 on CRC cells SW480 and SW837. (A) Axin2 protein levels increase, while active β-catenin and total β-catenin protein levels decreased after incubation with XAV-939 compared to the negative-control (DMSO) in SW480, and SW837 in a dose dependent manner. Proteins were isolated as whole protein lysates (left panel) and cytosolic and nuclear fractions (right panel). (B) Cellular viability was measured 48 hours after transfection using a CellTiter-Blue® assay. Exposure to XAV-939 significantly reduced cellular viability of SW837 but not SW480. (C) Cells where pre-incubated with XAV-939 and irradiated. Exposure to XAV-939 led to a significant radio sensitization of SW480 (1 μM P = 0.0116, 4 μM P = 0.0112; ANOVA model) and SW837 (5 μM P = 0.0223; 10 μM P = 0.000264; ANOVA model). The increased sensitivity to RT was stronger with higher doses of XAV-939. Each experiment was repeated three times. Data are displayed as mean values, n = 3, error bars {plus minus} SEM.

Supplementary Figure and Table Legends

Western blot analysis of (A) LS1034, (B) SW480, and (C) SW837 cells 24, 48, 72, and 96 hours after transfection with siRNAs targeting β-catenin, and a non-silencing control siRNA (siNEG).

Experimental conditions for cellular viability assays and dual luciferase reporter assays.

Supplementary Figure S8 PDF file - 391K, Reduction of CTNNB1 after siRNA against LNX2

Supplementary Figure S4 PDF file - 1138K, Cell viability and corresponding mRNA reduction of eight candidate genes selected for further experimentation

Supplementary Figure S12 PDF file - 1822K, Model for LNX2 mechanism of action

Supplementary Figure S9 PDF file - 1478K, Expression levels of the LNX2 module

Supplementary Figure S10 PDF file - 230K, Minimal region of amplification in 13q12.2

Supplementary Figure S11 PDF file - 415K, Overexpression of LNX2 in colorectal carcinoma compared to normal mucosa