Upregulation of nemo-like kinase is an independent prognostic factor in colorectal cancer

Figure 1 Representative immunohistochemical data of nemo-like kinase expression. Expression of NLK protein was mainly present in the cytoplasm of tumor cells, as indicated by the brown color (magnification × 200). CRC: Colorectal cancer.

Figure 2 Immunohistochemical detection of nemo-like kinase expression in familial adenomatous polyposis. Representative sections of A: Normal mucosa; B: Adenoma; and C: Adenocarcinoma; D: Individual; and E: Pooled quantification of staining from 16 familial adenomatous polyposis patients. ^bP < 0.01, ^dP < 0.001 via Wilcoxon test.

Figure 3 Immunohistochemical detection of nemo-like kinase expression in colorectal cancer. Representative sections of A: Normal mucosa; B: Primary tumor; C: Metastatic tumor; D: Individual; and E: Pooled quantification of staining from 21 patients with metastatic colorectal cancer. ^aP < 0.05 via Wilcoxon test.

Figure 4 Kaplan-Meier survival curves. A,C: Overall survival; B,D: Disease-free survival stratified by nemo-like kinase (NLK) expression in 712 colorectal cancer patients.

Figure 5 Knockdown of nemo-like kinase expression affects viability, colony formation, and migration of colorectal cancer cells. A: Nemo-like kinase (NLK) mRNA expression was assessed in six CRC cell lines using quantitative real-time-PCR; Knockdown of NLK B: mRNA and C: Protein expression in HT-29 cells 48 h after infection with a lentivirus carrying NLK (Lv-shNLK) or control (Lv-shCon) shRNA; D: Cell viability MTT assay; E: Colony formation assay at 14 d after infection; F: Transwell tumor cell migration assay. ^bP < 0.01, ^cP < 0.05; ^dP < 0.01; ^fP < 0.01.

Figure 6 Knockdown of nemo-like kinase expression affects cell cycle progression and matrix metalloproteinase-2 expression in HT-29 cells. A: Flow cytometric cell cycle distribution in HT-29 cells 48 h after infection with a lentivirus carrying nemo-like kinase (NLK) (Lv-shNLK) or control (Lv-shCon) shRNA; B: Summarized data of cell cycle distribution; matrix metalloproteinase (MMP)-2; C: mRNA; and D: Protein expression with NLK knockdown. ^bP < 0.01.