Conventional DCs from Male and Female Lupus-Prone B6.NZM Sle1/Sle2/Sle3 Mice Express an IFN Signature and Have a Higher Immunometabolism That Are Enhanced by Estrogen

Figure 2

Estrogen enhances the development of cDCs of B6 and TCSle mice of both sexes and the constitutive IFN signature in TCSle cDCs. Bone marrow precursors from B6 (black closed symbols) or TCSle (gray open symbols) female (circle) and male (triangle) mice were cultured with GM-CSF in standard phenol red/media conditions or media depleted of phenol red and devoid of steroids (charcoal-treated FBS: 0 E2) and supplemented with 0.03 nM, 0.1 nM, or 50 nM 17-β-estradiol (E2). On day 7-8, cDCs were harvested and stained with (a) antibodies against CD11c, CD11b, and (b) fixable viability dye and analyzed by flow cytometry or (c) total RNA was analyzed by qRT-PCR analysis. ISGs were normalized to the housekeeping gene cyclophilin. Standard condition female B6 was set to 1 in each experiment. Mean + SE values are from 6 independent experiments, using one mouse of each strain and sex per experiment. Two-way ANOVA analysis with Tukey multiple comparisons was used to calculate the significance of the effects of E2 treatment within each group of mice represented by brackets below the graph. A black star indicates statistical significance in all the fours curves representing both sexes and strains of cDCs. Two-way ANOVA analysis with Tukey multiple comparisons was used to compare differences between B6 and TCSle, and results are shown in a box surrounding the symbol Δ for significance between B6 and TCSle males or the symbol O for significance between B6 and TCSle females. , Δ, and O represent . ΔΔ, and OO represent . represent .

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