Extended Data Fig. 5: Spectral flow cytometry-based supervised analysis of the expression of tdTomato and GFP in splenocytes of ZeST mice.

a, Gating strategy followed to identify the indicated lymphoid vs myeloid cell populations. Splenocytes were isolated from uninfected or MCMV-infected ZeST mice. Cells were previously gated as CD45⁺ after exclusion of dead cells and doublets. b, c, The expression of CD11c (b), GFP (c) and of CD26 vs CD64 (d) was analyzed on Ly6C⁺ tDC cells (orange), Ly6C^neg tDC (purple) and CD26^neg Ly6C^neg cells gated as shown in (a). e, CD11c^high tDC (purple) and pDC-like cells (green) were gated from the spectral flow cytometry dataset according to the strategy used with the initial regular flow cytometry dataset (as shown in Fig. 3b), and then analyzed for CD26 vs Ly6C expression, for comparison with the gating strategy used to identify Ly6C^neg tDC and Ly6C⁺ tDC (panel a). f, Analysis of the expression of GFP vs the indicated cell markers upon gating on Ly6D^neg CX₃CR1^neg cells. g, Percentages (mean ± SD) of GFP⁺ cells (top) or tdT⁺ cells (bottom) in indicated cell populations gated as shown in (a). For (a) and (f), the analysis shown is representative one ZeST mouse out of 9 tested. For (b-e) n = 2 for NI, n = 4 for MCMV 36 h and n = 3 for MCMV 48 h.

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