Host control of metastasis in CZ, C3 and [CZ x C3] F1 mice.

<p>Host control of metastasis in CZ, C3 and [CZ x C3] F1 mice.</p

GD3 Supplementation of B4ST8 MEFs.

<p>GD3 Supplementation of B4ST8 MEFs.</p

Virus Internalization in B4St8 KO MEFs.

<p>B4St8 knock out MEFs were infected with MuPyV and then fixed at the indicated times post infection (30 min, 3 hrs). <b>(A)</b> Slides were stained for cell surface VP1 (red), and then permeabilized and stained for total VP1, showing both cell surface and intracellular VP1 (green). <b>(B)</b> At 30 mins post-infection line scan analysis shows that B4St8 MEFs exhibit similar staining for cell surface and total VP1, indicating minimal virus internalization. <b>(C)</b> At 3 hrs post-infection line scan analysis shows that B4St8 KO MEFs exhibit abundant intracellular VP1 staining (green only), indicating internalized virus.</p

Induction of c-fos in wild-type and B4St8 KO MEFs.

<p>Purified virus in serum-free medium was used to infect wild-type and B4St8 KO MEFs. Cell extracts were made at the indicated times post-infection and subjected to western blot analysis with anti-fos antibody.</p

IL-12 response to MuPyV by splenocytes in KO mice.

<p>IL-12 response to MuPyV by splenocytes in KO mice.</p

Susceptibility of wild-type and ganglioside deficient MEFs to small plaque and large plaque polyomavirus strains.

<p>Susceptibility of wild-type and ganglioside deficient MEFs to small plaque and large plaque polyomavirus strains.</p

Characterization of St8 and B4 KO mice.

<p>A. Kaplan-Meier survival curves for wild-type and ganglioside-knockout mice. Newborn mice were inoculated intraperitoneally with ~10⁶ PFU of the LID strain of virus and followed using death as an endpoint. B. Susceptibility of wild-type, ST8, and B4 MEFs to infection. Large T-antigen immunofluorescence of MuPyV-infected mouse embryo fibroblasts from wild-type and ganglioside-deficient mice. Cells were infected with the RA virus at an MOI of 1–2 PFU/cell.</p

Activation of NFAT and secretion of MMP-2 are critical for invasion by metastatic bone tumor cells.

<p>A) Immunoblot for NFAT c1 in tumor cell lysates. B) Normalized luciferase activity of an NFAT reporter in tumor cells. C) Inhibition of invasion by CZ II tumor cells. Left: cells ± TIMP-2. Middle: cells ± NFAT siRNA. Right: immunoblots on cells ± NFAT siRNA.</p

Metastatic tumor cells invade Matrigel via an MMP-2-mediated pathway.

<p>A) Migration assay was carried out as in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1000733#ppat-1000733-g002" target="_blank">Figure 2A</a> except the upper surface of the filter was coated with Matrigel. B) Upper – Zymography with gelatin as ‘in gel’ substrate and conditioned media from each of the tumor lines. Lower – Immunoblot for MMP-2 in tumor cell lysates.</p

Primary and metastatic bone tumors from CZ mice.

<p>A) A primary bone tumor (P) on the head of the femur of a CZ mouse. B) H&E stained section of the same tumor showing invasion of adjacent muscle and destruction of bone by invading tumor cells. C) Metastatic nodule (Mx) on the surface of the lung of the same animal. D) H&E stained section of the lung metastasis from the same animal showing osteiod. E) Occult metastatic lesion with osteoid in the lung of a different CZ animal. F) H&E stained section of a liver metastasis showing osteoid in a CZ animal inoculated with osteosarcoma cells.</p