Additional file 1: of High-performance gene expression and knockout tools using sleeping beauty transposon system

Figure S1. Re-expression of Flag-tagged RCC2 rescued sgRCC2-mediated delay of mitotic exit. A. HeLa cells expressing GFP-H2B were incubated with Nocodazole (100 ng/ml) or Taxol (2 μM) for 12 h, and analyzed using ZEISS710 confocal microscope. (Scale bars, 2 μm). B. Immunoblots for control cells, RCC2-depleted cells, and RCC2-depleted cells stably expressing the indicated constructs. C. Time-lapse images showing mitotic exit in HeLa-H2B cells indicated above. Figure S2. Depletion of BRD7 delays mitotic exit and leads to spindle misorientation in HeLa cells. A. HeLa cells transfected with indicated sgRNA and synchronized in M phase by incubation with 100 ng/ml nocodazole for 14 h. M phase cells selected by shake-off were released for the indicated time. B. Time-lapse images showing prolonged metaphase to anaphase and misoriented cell division (uneven timing of daughter-cell adhesion to the substratum) in BRD7-depleted HeLa-H2B cells, compared with control. (Scale bars, 2 μm). (DOCX 7450 kb

DYRK2 inhibits the proliferation, cell migration and invasion of colorectal cancer cells in vitro.

A, B: The effect of DYRK2 depletion on the cell cycle. Experiments were performed in triplicate. C: Western blot analysis of E-cadherin, N-cadherin, Fibronectin and Vimentin expression in the indicated treatment of the CRC cell lines. D, E: The cell's ability of wound gap closure was dramatically enhanced by knocking down DYRK2 both in HCT116 and SW480 cells. F, G: Migratory and invasive abilities of the indicated cell lines in vitro was measured by the Transwell assay as described in “Materials and methods”. Bars correspond to mean+ standard error, with a P value calculated by using Student’s t-test. *PP<0.001.</p

Clinicopathological characteristics and DYRK2 expression of 181 patient samples of CRC.

<p>Clinicopathological characteristics and DYRK2 expression of 181 patient samples of CRC.</p

Decreased expression of DYRK2 mRNA in colorectal cancer tissues.

<p>Real-time PCR analysis of DYRK2 mRNA expression in sixteen primary colon cancer tissues (T) and adjacent non-cancerous tissues (ANT) paired from the same patient.</p

The clinicopathologic features of the 16 CRC samples.

<p>The clinicopathologic features of the 16 CRC samples.</p

The detail information of the six Oncomine datasets.

<p>The detail information of the six Oncomine datasets.</p

mRNA expression of DYRK2 is lower in high grade and metastatic CRC compared to low grade and non-metastatic CRC.

A. Bittner's colorectal carcinoma data set: low grade CRC vs high grade CRC; B. Bittner's colorectal carcinoma data set: non-metastatic CRC vs metastatic CRC; C. Ki's colorectal carcinoma data set: adjacent normal colon vs primary CRC without liver metastatic CRC vs primary CRC with liver metastatic CRC and secondary CRC metastasis at liver; D. Ki's colorectal carcinoma data set: paired primary CRC with liver metastatic CRC vs paired secondary CRC metastasis at liver.</p

Kaplan-Meier curves with univariate analyses (log-rank) for patients with low DYRK2 expression (dotted line) versus high DYRK2 expression (bold line).

A, Overall survival of patients (clinical stages I-IV) with either low or high DYRK2 expression. B, Overall survival of patients (clinical stages I-II) with either low or high DYRK2 expression. C-D, Overall survival of patients (clinical stages III (C) and IV (D)) with either low or high DYRK2 expression.</p

Univariate and multivariate analysis of various prognostic parameters in patients with CRC Cox-regression analysis.

<p>Univariate and multivariate analysis of various prognostic parameters in patients with CRC Cox-regression analysis.</p

mRNA expression of DYRK2 is lower in CRC compared to the normal colon/rectum.

<p>A. Gaedcke's colorectal carcinoma data set: paired normal colon vs paired CRC; B. Skrzypczak's colorectal carcinoma data set: Normal colon vs colorectal adenocarcinoma and colorectal carcinoma; C. Kaiser's colorectal carcinoma data set: normal colon vs different types of CRC; D. TCGA colorectal carcinoma data set: normal colon and rectum vs different type of CRC.</p