Figure 2. MLKL signaling does not influence Langat virus pathogenesis.

(A-B) Survival analysis (A) and presentation of clinical signs of disease (B) in Mlkl^−/− mice and littermate controls following subcutaneous inoculation with 3×10⁴ PFU LGTV TP21. Data are pooled from two experiments. (C) Mlkl^−/− and littermate control mice were infected subcutaneously with LGTV TP21. On indicated days following infection, splenic viral burden was measured via qRT-PCR. Data was normalized against a standard curve of known viral titers to generate plaque-forming unit (PFU) equivalents. Data for each day post infection are pooled from 2–3 experiments. LOD, limit of detection. (D-P) Mlkl^−/− and littermate control mice were infected subcutaneously with LGTV TP21 for 8 days prior to harvesting splenocytes and profiling leukocytes by flow cytometry. (D) Representative flow cytometry plots showing CD8+ and CD4+ T cells among CD3+ leukocytes in the spleen. Numbers represent percentage of cells in each gate relative to total plotted cells. (E-F) Numbers of CD8+ T cells (E) and CD4+ T cells (F) among CD3+ leukocytes. (G-H) Percentage of CD44+ cells among CD8+ T cells (G) and CD4+ T cells (H). (I-N) Numbers of CD19+ B cells (I), CD11b+ NK1.1+ Natural Killer cells (J), CD11c+ MHC-II+ dendritic cells (K), CD45high CD11b+ F4/80+ macrophages (L), CD11b+ Ly6G+ neutrophils (M), and CD45high CD11b+ Ly6C+ monocytes (N) among total leukocytes in the spleen. (O-P) Percentage of CD80+ cells among CD11c+ MHC-II+ dendritic cells (O) and CD11b+ F4/80+ macrophages (P). ns, not significant. Error bars represent SEM.