Supplementary Figure 8: Gating strategy for AM and MO subsets in BM, blood and BALF following infection with MuHV-4.

(a-c) Flow plots on MO subsets in BM, gated as liveCD19^-CD11b⁺SiglecF-Ly6G^-Ly6C⁺ cells (a), on MO subsets in blood gated as live CD11b⁺Ly6G^-CD19^-SiglecF^-Ly6C⁺ cells (b), on AM defined as liveautofluorescent⁺CD11c^hi cells (c) and BALF MO gated as nonautofluorescentCD11c^loCD19^-CD11b⁺Ly6G^-CCR2⁺Ly6C⁺ further analyzed for MHCII and Sca-1 expression (c). (d) Representative flow cytometry overlays of AM (as defined above) from BALB/c mock-infected mice (red) and AM (green) and MOs (blue) from MuHV-4 infected mice isolated from BALF at the different times p.i.. (e,f) Flow cytometry quantification of AM viability in BALF at different times p.i. using Annexin V-APC/7-AAD staining. Representative flow cytometry plots (e) and quantification of Annexin⁺/7AAD^- and Annexin⁺/7AAD⁺ cells among AM (f) from mock or MuHV-4-infected BALB/c mice at different times p.i.. Data are mean ± s.e.m. of samples from 5 mice per group. *** p< 0.001, ** p< 0.01 and * p < 0.05 (in (f), all data were compared to values obtained at day 0 by one-way ANOVA and Dunnett's multiple comparison test).