IHC analysis of MRGD expression.

<p>Cell block samples were used to confirm the antibody specificity in immunostaining. HEK293/αvβ3 cells transfected with MRGD expression vector (<b>A</b>) or Mock vector (<b>B</b>) are shown. <b>C.</b> The representative staining of lung adenocarcinoma which is positive for MRGD.</p

Tumorigenesis of NIH3T3 cells stably expressing MRGD (NIH3T3-MRGD cells).

<p><b>A.</b> Representative pictures of focus formation in monolayer cultures of NIH3T3 cells stably expressing Mock (NIH3T3-Mock cells, left) or MRGD (NIH3T3-MRGD cells, right). Cells were stained with crystal violet after fixing with 4% paraformaldehyde. <b>B.</b> Representative pictures of NIH3T3-MRGD or NIH3T3-Mock spheroid on Days 1 and 7. <b>C.</b> Spheroid growth curves. The spheroid sizes of NIH3T3-MRGD (closed) or NIH3T3-Mock (open) at Days 2, 3, 5 and 7, are shown with their diameters (mean ± SD). * indicates p<0.005 (Mann-Whitney U test, 2 tails). <b>D.</b> Cell proliferation of spheroid cultures of NIH3T3-MRGD or NIH3T3-Mock. Luminescence of whole cell ATP contents (means ± SD) was measured at 6 days after plating. * indicates p<0.005 (Mann-Whitney U test, 2 tails).</p

MRGD ligand, beta-alanine, promoted cell proliferation of NIH3T3-MRGD cells.

<p>NIH3T3 cells transfected with MRGD or RASV12 were cultured in RPMI1640 containing 0.1% BSA and various concentration of beta-alanine for 7 days. Data were obtained from three independent cell cultures (means ± SD). * indicates p<0.05 (Mann-Whitney U test, 2 tails), compared with NIH3T3-MRGD without beta-alanine, and with NIH3T3-RASV12.</p

Tumor volumes of NIH3T3-MRGD or Mock bearing mice.

<p>Tumor volumes (mm³) were calculated according to the following equations: Tumor volume (mm³) = 1/2×(tumor length)×(tumor width)²</p

Quantitative RT-PCR analysis of MRGD mRNA expression in clinical samples.

<p>One hundred and twenty seven sets of RNA samples of cancer and non-cancer portions from the same patients with lung (n = 33), esophagus (n = 12), breast (n = 16), kidney (n = 10), stomach (n = 25), uterus (n = 12) or colon (n = 19) cancer. Ratios of the amount of MRGD mRNA in tumor portion per that in normal portion of each case were plotted. The bar indicates the mean of the ratio in each cancer type.</p