DSA imaging of treated tumors.

<p>The tumors showed hypervascularity in the liver as determined with DSA imaging (A, black arrow). After lipiodol (60°C) injection, the tumors are completely or largely de-vascularized and show on DSA as a lipiodol-filling defect (B, black arrow).</p

TEM results of treated tumors.

<p>Stimulated by heated (60°C) lipiodol perfusion, TEM results revealed that the tumor endothelial cell microvilli decreased (A, 5000×), the vascular endothelial cell endoplasmic reticulum expanded, and the chondriosome was swollen (B, 10000×).</p

Comparisons of PCNA protein levels between groups after perfusion.

<p>PCNA: control <i>vs.</i> treated group, <i>P</i> = 0.047.</p

Expression of VEGFR and VEGF protein and mRNA levels.

<p>The relative changes in VEGFR or VEGF protein and mRNA levels in tumor tissue were detected after treatment in each group (n = 10). A and B. VEGFR and VEGF mRNA expression levels were evaluated using real-time quantitative PCR as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061583#s2" target="_blank">Materials and Methods</a> section. C and D. VEGFR and VEGF protein levels were detected using Western blot analysis (upper panel). β-actin was detected as a loading control. VEGFR and VEGF expression levels were quantified through densitometry and plotted as the fold change (lower panel). The values are presented as the mean±SD of 3 independent experiments (* <i>P</i><0.05 <i>vs.</i> control group).</p

Expression of PCNA protein.

<p>As detected through immunohistochemistry, PCNA protein expression was detected mainly in viable VX2 tumor cells (brown; A, control 400<b>×</b>; B, treated 400<b>×</b>).</p