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 7

Estrogen enhances CXCL10 chemokine production not predicted by Cxcl10 RNA. Bone marrow precursors from B6 (black closed symbols) or TCSle (gray open symbols) female (circle) and male (triangle) mice were cultured in standard conditions or hormone-depleted conditions supplemented with 0.03 nM, 0.1 nM, or 50 nM E2 (a, b, e, f). TCSle cDCs were cultured in standard conditions supplemented with Fulvestrant (Fulvi 1 μM or 100 nM in DMSO) and Tamoxifen (Tam 10 nM or 100 nM in ethanol) (c, d). On day 7, cDCs were stimulated with CpG (10 μg/mL) (a, c, e) or R848 (1 μg/mL) (b, d, f). Supernatants were harvested 24 hours post stimulation and analyzed by ELISA for CXCL10 protein levels (a–d). Total RNA was isolated 6 hours post stimulation and analyzed by qRT-PCR (e, f). Mean + SE values are from 3 (female cDCs) or 4 (male cDCs) independent experiments or (c and d) 2 independent experiments using one mouse per strain per experiment (a, b, e, f). 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. Black indicates significance within all 4 groups while symbols represent significance within a single group. 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 .

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