FIG 2.

Analysis of FV-mWasabi-infected cells. C57BL/6 mice were infected with 20,000 SFFU FV-mWasabi, and infected target cells were analyzed in bone marrow, lymph nodes, and spleens on days 2, 4, 7, 14, 31, and 42 after FV-mWasabi infection. For the identification of infected cell types, isolated cells were stained with a target cell antibody panel and analyzed by flow cytometry. Minimal manual gating was performed to gate on singlets, live cells, and mWasabi⁺ cells, which were concatenated and subjected to a t-SNE analysis. (A) Manual gates on samples from individual organs and days of analysis were overlaid on the t-SNE plot to visualize the samples. (B) Manual gates on individual cell markers were overlaid on the t-SNE plot to identify cell types. (C and D) Manual gating was performed to determine the number of mWasabi⁺ cells in Ter119⁺ cells (ery), Gr1⁻ CD11b⁺ CD11c⁺ myeloid cells (Gr1⁻ My), Gr1⁺ CD11b⁺ myeloid cells (Gr1⁺ My), CD19⁺ B220⁺ B cells, and CD4⁺ or CD8⁺ CD3⁺ T cells, which is shown as the number of mWasabi⁺ cells in 10⁶ cells (C) and as the frequency of these cell types in the total population of mWasabi⁺ cells (D). Each circle represents the value for an individual mouse, and bars depict mean values for groups of mice. The dotted line indicates the detection limit. The data for each time point were obtained from two (day 2, day 4, day 14, and day 31), three (day 7), or six (day 42) independent experiments (n = 8 [day 2], 9 [day 4], 10 [day 7], 8 [day 14], 9 [day 31], or 28 [day 42]). Statistically significant differences (P < 0.05) between the number (C) or frequency (D) of infected cells of the different organs within a cell subset are indicated by lines and asterisks; color-coded dagger or double-dagger symbols indicate dominant subsets with a significantly higher number (C) or frequency (D) of FV-mWasabi-infected cells compared to three (dagger) or four (double dagger) other subsets within one organ.