The overexpression of Cry1 mRNA and protein in colorectal cancer tissues.

<p>(<b>A</b>) A representative image of Cry1 staining in colorectal cancer tissues is shown. (<b>B</b>) A representative image of Cry1 staining in adjacent noncancerous tissues is shown. (<b>C</b>) Cry1 protein expression level was higher in tumor tissues compared to adjacent control tissue as detected by immunoblotting (mean±SEM; n = 109; * **, <i>P</i><0.001). (<b>D</b>) Average T/N ratios of Cry1 mRNA expression in paired colorectal cancer (T) and normalmucosa tissues (N) were quantified by qPCR and normalized to GAPDH. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments. Magnification is ×200.</p

The expression of Cry1 protein in colorectal cancer sections.

<p>Representative immunohistochemical images of colorectal cancer tissue specimens indicating negative or weakly detectable Cry1 staining (<b>A</b> and <b>B</b>); moderate Cry1 staining (<b>C</b>); and strong Cry1 staining (<b>D</b>) are shown. Magnification is ×200 (<b>A, B, C</b> and <b>D</b>).</p

Overexpression of Cry1 mRNA and protein in colorectal cancer cell lines.

<p>Expression of Cry1 mRNA and protein in colorectal cancer cell lines (SW480, SW620, HT29, THC8307, and HCT116) and FHC were examined by qPCR (<b>A</b>) and Western blotting (<b>B</b>). Expression levels were normalized to GAPDH. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments, *, <i>P</i><0.05.</p

Cry1 promoted CRC growth <i>in vivo</i>.

<p>(<b>A</b>) The expression of Cry1 was markedly increased in the stable cell line Cry1-HCT116 compared with control stable cell line ctrl-HCT116. GAPDH was used as an internal control. (<b>B</b>) Representative images of tumors derived from Cry1-HCT116 or ctrl-HCT116,following subcutaneous xenograft transplants in nude mice (<b>C</b>) Overexpression of Cry1 promoted colorectal cancer growth. Tumor cells were injected subcutaneously into nude mice. Mice were sacrificed after 4 weeks, and the volume of each tumor was measured every 4 days. Bars, ±SEM; *<i>P</i><0.05, ** <i>p</i> = 0.01.</p

Effects of Cry1 on cell growth.

<p>(<b>A</b>) Cry1 protein levels were upregulated in HCT116 cells and downregulated in SW480 cells. (<b>B</b>) MTT assays on HCT116 cells 24, 48, and 72 h after transfection with Cry1 or GFP control (***, p<0.001). (<b>C–D</b>) Colony formation assay of HCT116 cells transfected with Cry1 or GFP control (*, <i>p</i> = 0.033). (<b>E–F</b>) Inhibition of SW480 cell colony formation capacity by Cry1 siRNA relative to control (**, <i>p</i> = 0.007). Experiments were repeated at least three times, and representative data are presented; <i>bars</i>, SD.*, <i>P</i><0.05; **, <i>P</i><0.01, ***, <i>p</i><0.001.</p

Clinicopathological findings and correlation with Cry1 expression.

*<p>Statistically significant. Numbers in parentheses indicate the proportion of tumors with a specific clinical or pathological feature in a given Cry1 subtype.</p>†<p>Analysis for this parameter was available for 167 cases.</p>††<p>Analysis for this parameter was available for 149 cases.</p>†††<p>Analysis for this parameter was available for 147cases.</p

The level of Cry1 protein expression affects overall survival and disease-free survival.

<p>Kaplan–Meier curves with univariate analysis (log-rank) for colorectal cancer patients with high Cry1 expression (n = 101) versus low or no Cry1 expression for overall survival (n = 67) (<b>A</b>) and disease-free survival (n = 67) (<b>B</b>) are shown. Higher expression of Cry1 positively correlated with the poor patientoutcomes.</p

MOESM1 of Synthetic lethal short hairpin RNA screening reveals that ring finger protein 183 confers resistance to trametinib in colorectal cancer cells

Additional file 1: Table S1. Abundance of genomic short hairpin RNA (shRNA) fragments under different conditions