A shows a volume rendering of a mouse 30 minutes after i.v. injection of ExiTron nano 12000.

<p>B is a curved maximum intensity projection in coronal orientation of the same scan. A and B demonstrate the feasibility to perform CT angiography during the early intravascular phase of the tested contrast agent. Additionally, A and B show the early contrast agent uptake by the RES with increasing contrast of liver and spleen. C is a coronally oriented curved maximum intensity projection of a mouse that did not receive contrast agent.</p

A and B show intrasplenic (*) and intrahepatic (LMet) growing tumors 26 days after intrasplenic injection of C15A3 colon tumor cells.

<p>A and B were acquired 4 hours after i.v. injection of 100 µl ExiTron nano 12000. B, C, and D illustrate contrast enhancement of the abdominal and mediastinal lymph nodes (LN) and of the adrenal glands (AdrG). C was acquired 4 hours after i.v. injection of 100 µl ExiTron nano 12000; D was acquired 22 days after i.v. injection of 100 µl ExiTron nano 12000. Micro-CT scanning parameters: 40 sec scan time; 190° rotation; 1200 projections; voxel size 41×41×55 µm³.</p

Transcript levels, protein levels and proteolytic activity of Separase in BCR-ABL-positive cells treated with IM.

<p>Cells were treated individually with distinct concentrations (0.5 to 5 µM) of IM. After about two cell cycle rounds (K562, LAMA-84, 24 h; U937p210BCR-ABL/c6-On, 48 h) total RNA and protein lysates were prepared and analyzed by qRT-PCR (A), Western blot immunostaining (B) and separase fluorometric activity assays (C). For Western blot experiments, Actin served as loading control and/or for densitometric data normalization. Each data point corresponds to one single experiment. Only significant p-values as calculated between treated and untreated cells were shown (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042863#pone-0042863-t002" target="_blank">Table 2</a> for summarized Δ-values). For a representative set of corresponding immunostained Western blots compare <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042863#pone-0042863-g005" target="_blank">Figure 5</a> panel C. Abbreviations: U937/c6-On, U937 cells expressing a p210BCR-ABL transgene (Tet-On system) after induction with Doxycycline (U937p210BCR-ABL/c6-On).</p

Time course of contrast enhancement within the vascular system and the liver of C57BL/6J mice (n = 3 per group) after a single i.v. injection of 100 µl ExiTron nano 6000 or ExiTron nano 12000.

<p>Measurements were performed by placing a ROI within the left ventricle (vessel contrast) and within the liver avoiding large intrahepatic vessels. The baseline level ( = 100%) refers to measurement of the relative density of the liver and the vascular system prior to administration of contrast agent.</p

Axial reconstructed micro-CT images.

<p>Micro-CT of the liver of a C57BL/6J mouse was performed 3 hours after injection of a nanoparticular contrast agent (ExiTron nano). The color-coding of the single liver lobes corrsponds to the color coding presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031179#pone-0031179-g001" target="_blank">Figure 1</a>. The orientation of the axial slices in relation to the spine is shown in the lower right corner.</p

Liver variationes.

<p><b>A–C</b> are comparable coronal sections of the murine liver illustrating the variability of the size of the papillary process of the caudate lobe. <b>D</b> shows transdiaphragmatic herniation of parts of the right medial liver lobe, which we incidentally discovered during our studies.</p

A and B show a partial diaphragmatic herniation of the left upper liver lobe in coronally (A) and sagittally (B) reconstructed maximum intensity projections of a C57BL/6J mouse 22 hours after i.v. injection of 100 µl ExiTron nano 12000.

<p>The herniated liver tissue can be easily delimited from the adjacent heart due to the positive liver contrast. C and D are micro-CT scans of the murine liver before (C) and 24 hours after (D) intravenous administration of 100 µl ExiTron nano 6000.</p

Illustrative micro-CT images of liver metatstases in a mouse.

<p><b>A–C</b> Images of liver metastases were acquired 2 hours after i.v. injection of 100 µl of a contrast agent (Viscover Exitron nano 6000; Miltenyi Biotec, Bergisch-Gladbach, Germany) in a coronal (A), transversal (B), and saggital (C) slice orientation. With regard to liver anatomy, we found metastases to develop predominantly under the liver capsule adjacent to the fissures between the liver lobes. While the left lateral lobe (LLL) shows no metastases on its caudal edge in this slice, there are plenty on its cranial edge, next to the left medial lobe (LML). The lobes on the right side (right medial lobe (RML), right lateral lobe (RLL) and the caudate process (CP)) show (besides the subcapsular growth pattern) metastases within the liver parenchyma. Due to tumor growth the papillary process depicted in B and C lost its typical shape (compare with <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031179#pone-0031179-g002" target="_blank">figs. 2</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031179#pone-0031179-g003" target="_blank">3</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031179#pone-0031179-g004" target="_blank">4</a>).</p

Hepatic veins.

<p>Maximum intensity projection (MIP) of micro-CT images of the murine hepatic veins 20 minutes after injection of 400 µl Fenestra VC. The color coding of the draining veins corresponds to the color coding used for the liver lobes before. The renal veins are indicated by arrowheads.</p

Analysis of master Separase proteolytic activity regulators in BCR-ABL-negative and -positive cell lines treated with IM.

<p>Schematic diagram of cooperating inhibitory factors that regulate Separase proteolytic activity in a tight cell cycle controlled manner (<b>A</b>). Degradation of Securin, inactivation of the CyclinB1/CDK1 complex, dephosphorylation of Separase at a specific serine residue (pSer1126) by the anaphase promoting complex/cyclosome (APC/C), and the release of PP2A contribute to activation of Separase. Representative composite image of Western blot immunostaining experiments illustrate the expression levels of Separase and relevant regulatory proteins (Securin, pSer1126, CyclinB1 and PP2A) in BCR-ABL-negative (<b>B</b>) and BCR-ABL-positive (<b>C</b>) cell lines. Images are cropped sections derived from stripped and reprobed Western blot immunostainings used for acquisition of densitometric data shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042863#pone-0042863-g002" target="_blank">Figures 2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042863#pone-0042863-g004" target="_blank">4</a>. Cells were treated with IM for times and doses given on top. Actin served as loading control. The densitometric data of at least triplicate experiments are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0042863#pone-0042863-t003" target="_blank">Table 3</a>.</p