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 10

E2 enhances the higher energy metabolism of TCSle cDCs. cDCs from B6 (darker color) or TCSle (lighter color) female (red/orange) and male (blue/azur) mice were cultured in standard conditions or in hormone-depleted conditions supplemented with 0.03 nM, 0.1 nM, or 50 nM E2 (a–k). On day 7, cDC were stimulated with CpG B 1826 (10 μg/mL) or R848 (1 μg/mL). Supernatants were harvested and analyzed using the Griess reaction 24 hours after stimulation (a–e). Six hours post stimulation, cDCs were harvested for qRT-PCR analysis (f–j). Nos2 (f–j) and Pdk1 (k) genes were normalized to the housekeeping gene cyclophilin. Standard female B6 condition was set to 1. Mean + SE values are from 3 independent experiments, using one mouse per strain per experiment. Two-way ANOVA analysis with Tukey multiple comparisons was used to determine the significant activation by CpG or R848 within each group of mice, represented by brackets below the graph. Black indicates significance within all 4 groups while individual colors/symbols represent significance within a single group (a–k). Two-way ANOVA analysis with Tukey multiple comparisons, used to compare differences between females and males, is represented by brackets and above the graph (a–e). Tukey multiple comparisons, used to compare differences between B6 and TCSle, is represented by a box surrounding red Δ symbol for B6 and TCSle females or a blue O symbol for B6 and TCSle males (a–j). Two-way ANOVA analysis with Tukey multiple comparisons is used to determine the effects of E2 treatment on cDC differentiation within each group of mice represented by brackets and symbols below the graph (j). , Δ, and O represent . , ΔΔ, and OO represent . represents

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