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. 2011 Aug 25;7(8):e1002157. doi: 10.1371/journal.ppat.1002157

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Wang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Confocal microscopy of pOka- or pOka-mSIM-N&C-infected skin cells that were stained for ORF61 (red), PML (green), and nuclei (blue). The images show association of ORF61-positive puncta with PML NBs in newly infected cells. Scale bars: 2.5 µm. (B) Quantitation of ORF61/PML NB association in pOka- or pOka-mSIM-N&C-infected cells that had ORF61-positive puncta. The left panel shows percentage of cells that show/did not show ORF61 colocalization with PML NBs (pOka, N = 47; mSIM-N&C, N = 46; ***, p<0.0001 by Chi-square). The right panel shows percentage of ORF61-positive puncta colocalized/not colocalized with PML NBs (pOka, N = 167; mSIM-N&C, N = 202; ***, p<0.0001 by Chi-square). (C) Confocal microscopy of pOka- or pOka-mSIM-N&C-infected skin cells that were stained for ORF23 (red), PML (green), and nuclei (blue). For each panel (left and right), the upper left image shows skin structures of a parallel section by hematoxylin and eosin (H+E) staining; the upper right image shows epidermal cells from the white square area on the upper left image (the infected cells were identified by ORF23 expression); the lower images show PML NBs in ORF23-positive cells and neighboring uninfected cells from the white square area on the upper right image. White arrows indicate cells with abundant nuclear ORF23 protein. Scale bars: 10 µm. (D) PML NB frequencies (mean±SEM) in pOka- or ORF61 SIM mutants-infected skin cells (pOka, N = 153; mSIM-C, N = 143; mSIM-N&C, N = 108; ***, p<0.0001 by t-test). (E) PML NB frequencies (mean±SEM) in uninfected skin cells from pOka- or pOka-mSIM-N&C-infected skin xenografts. Cells from the three major epidermal layers were grouped. Uninfected cells from pOka-infected skin: basal layer cells (N = 99), spinous layer cells (N = 79), granular layer cells (N = 45). Uninfected cells from pOka-mSIM-N&C-infected skin: basal layer cells (N = 109), spinous layer cells (N = 124), granular layer cells (N = 39).