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. 2014 Aug 14;10(8):e1004326. doi: 10.1371/journal.ppat.1004326

Figure 9. EVM005 inhibits activation of the immune response.

Figure 9

C57BL/6 mice were infected with 10,000 pfu of ECTV or ECTV pfu of ECTV or ECTV-Δ005 via intranasal inoculation. Tissues and whole blood were collected at 2, 3, 4 and 7 days post infection. (A and B) Spleen, liver, lung and kidney tissues were homogenized and plated onto BSC-1 cells to measure viral titers at day 4 (A) and 7 (B) post infection. Cell suspensions were prepared from infected spleens (C and D) or peripheral blood (E and F) on day 3 and 7 and analyzed by flow cytometry. (C and E) NK cells are identified as CD45+, CD3−, and NK1.1+. We gated 50,000 CD45+ cells, and presented the total number of NK1.1+ cells within this lymphocyte population (D and F) Virus-specific CD8+ T-cells were identified as CD8+, CD45+, and tetramer+, and presented as the ratio of tetramer+ cells to total CD8+ T-cells within each sample. (G and H) RNA was purified from spleen (G) and liver (H) and subjected to qRT-PCR to quantify transcriptional upregulation of TNFα, IL-1β and IL-6 at 7 days post infection. Data are normalized against GAPDH.