The three blood samples containing CTM from liver, nasopharyngeal and breast cancers.

<p>The three blood samples containing CTM from liver, nasopharyngeal and breast cancers.</p

CTCs detected in a blood sample from a liver cancer patient.

<p>A total of 10 CTCs were detected in this sample; 3 single migratory biophenotypic epithelial/mesenchymal CTCs, 3 single migratory mesenchymal CTCs and a tumor microembolus containing 4 mesenchymal CTCs were observed (epithelial biomarkers are indicated by red fluorescence; mesenchymal biomarkers are indicated by green fluorescence).</p

Calibration curve obtained using the optimized CanPatrol CTC enrichment technique in the spiking experiment (n = 8) using HepG2 cells at different dilutions.

<p>Calibration curve obtained using the optimized CanPatrol CTC enrichment technique in the spiking experiment (n = 8) using HepG2 cells at different dilutions.</p

Information and clinical characteristics of the patients.

<p>Information and clinical characteristics of the patients.</p

EpCAM, CK8/18/19, vimentin and twist expression in HepG2 tumor cells and leukocytes.

<p><b>A</b>: negative control, leukocytes stained for CD45 expression (bright blue fluorescence); <b>B</b>: HepG2 cells stained for EpCAM expression (red fluorescence); <b>C</b>: HepG2 cells stained for CK8 expression(red fluorescence); <b>D</b>: HepG2 cells stained for CK18 expression(red fluorescence); <b>E</b>: HepG2 cells stained for CK19 expression(red fluorescence); <b>F</b>: HepG2 cells stained for vimentin expression (green fluorescence); <b>G:</b> HepG2 cells stained for twist expression(green fluorescence); <b>H:</b> HepG2 cells stained for EpCAM, CK8/18/19, vimentin and twist expression (red/green fluorescence). The cells were analyzed using a 100x oil objective</p

The Kaplan-Meier survival curves of patients with positive TRIM24 expression and negative TRIM24 expression.

<p>The Kaplan-Meier survival curves of patients with positive TRIM24 expression and negative TRIM24 expression.</p

Depletion of TRIM24 reduces cell proliferation.

<p>4(A). Western blotting analysis of the cell-cycle related proteins showed the expression of Cyclin D1 and CDK4 were decreased but showed no significant change in p21 after knockdown TRIM24 in HepG2 cells; 4(B). Cell cycle analyses showed that the percentage of G1 phase was increased in siTRIM24 group (P<0.05), whereas the percentages of S phase (P<0.05) and G2 phase (P<0.05) were decreased in the TRIM24 knockdown cells compared with control cells; 4(C) CCK-8 assay suggested that cell proliferation of HepG2 after TRIM24 silencing was reduced compared with the control group.</p

Immunohistochemical staining of TRIM24 in tissue sections.

<p>A. Negative staining in normal liver tissue. B. Negative staining in benign liver lesions tissues (Hepatic hemangioma). C. Negative TRIM24 staining in an AFP>400 ug/L, well differentiated HCC tissue. D. Positive TRIM24 staining in an AFP<400 ug/L, moderate differentiated HCC tissue. E. Negative TRIM24 staining in an AFP<400 ug/L,well differentiated HCC tissue. F&G. Positive TRIM24 staining in an AFP>400 ug/L, poor differentiated HCC tissue. H. Negative control using antibody diluent.</p

Relationship between the TRIM24 expression and the clinicopathological factors.

<p><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0085462#pone-0085462-t001" target="_blank">Table 1:</a> a. Missing data for 2 patients. b. Missing data for 3 patients. c. Missing data for 1 patient. d. Missing data for 10 patients; 7 patients were excluded according to the inclusion criteria. *.We defined that those patients whose recurrence time is less than 6 months as intrahepatic metastasis.</p

Depletion of TRIM24 inhibits the process of EMT.

<p>5(A). Western blotting analysis of the cell-apoptosis related proteins showed the expression of E-cadherin was increased and Snail, Slug, Vimentin, andβ-catenin were decreased after knockdown TRIM24 in HepG2 cells; 5(B). Reduced migration and invasion ability of HepG2 cells at 48 h post-transfection with siTRIM24 (P<0.05, compared with controls).</p