Expression of IFN-I and ISGs in infected NFAT5-deficient macrophages and DCs. (A and B) Gating strategy for sorting spleen pDCs (A) and bone marrow pDCs and macrophages (B) after in vivo acute LCMV infection (2 × 10² PFU/mouse). (C and D) mRNA expression of the indicated genes in bone marrow macrophages (C) and pDCs (D) 3 d after infection with LCMV (n = 3 wild-type Nfat5^+/+ Vav-Cre mice and n = 5 Nfat5^fl/fl Vav-Cre mice). *, P < 0.05. (E) Percentage of bone marrow and spleen populations 3 d after LCMV infection (n = nine mice of each genotype for bone marrow populations and six for spleen, analyzed in two independent experiments). N, Ly6G⁺ neutrophils; M, CD11b⁺ F4/80⁺ macrophages. (F) Wild-type (Nfat5^+/+) and NFAT5-deficient (Nfat5^−/−) BMDCs differentiated with Flt3L and sorted as conventional DCs (CD11c⁺, B220⁻) were infected with VSV (top) or LCMV (bottom). mRNAs of IFN-I genes and Ifit2 were analyzed by RT-qPCR. Results show the mean ± SEM of six BMDC cultures of each genotype for VSV and four for LCMV analyzed in four independent experiments. mRNA values for each gene were normalized to L32 RNA and are represented relative to infected wild-type cells. (G) Relative mRNA levels for IFN-I genes and ISGs Mx1 and Ifit2 in wild-type and NFAT5-deficient BMDMs either not infected (n.i.) or after infection with VSV-GFP (MOI 1, 12 h) and isolating GFP⁺ and GFP⁻ cells by FACS sorting. Results show the mean ± SEM of three independent experiments. mRNA values for each gene were normalized to L32 RNA and are represented relative to the GFP⁻ wild-type cells. Statistical significance in C and D was determined with an unpaired t test, in F with a one-sample t test using infected wild-type cells as reference, and in G with a one-sample t test for comparisons with GFP⁻-sorted wild-type BMDMs or with an unpaired t test between the other samples. *, P < 0.05; **, P < 0.01.