GD3 Supplementation of B4ST8 MEFs.

<p>GD3 Supplementation of B4ST8 MEFs.</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

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

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

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

Virus Internalization in Wild-Type MEFs.

<p>Wild-type MEFs were infected with MuPyV and then fixed at the indicated times post infection (30 min, 3hrs). <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 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 MEFs exhibit abundant intracellular VP1 staining (green only), indicating internalized virus.</p