Hy 12870

Four weeks after treatment, 60 rats (n = 20 per group) were randomly selected and equally divided into ERα group and ERβ group. Five rats were randomly selected from the SHAM group and OVX + E group of ERα group, the antagonists of ERα (AZD9496, HY-12870, MedChemExpress) were injected into POA, and the agonists of ERα (Propyl pyrazole triol, HY-100689, MedChemExpress) were injected into POA of the five rats randomly selected from the OVX group. Five rats were randomly selected from SHAM group and OVX + E group of ERβ group to inject antagonists of ERβ (PHTPP, HY-103456, MedChemExpress) into POA, and five rats from OVX group were randomly selected to inject agonists of ERβ (Liquiritigenin, HY-N0377, MedChemExpress) into POA. Other rats were given the same volume of cosolvent (10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline, MedChemExpress) in POA. After 72 h of drug injection, the POA was removed according to the above method, and the mRNA and protein expression of Vglut2 and Vgat in POA were detected by Western blot and qRT-PCR, respectively. The details on the primary and secondary antibodies of vglut2 and vgat are shown in Supplementary Table 1.1, and the details on the primers of vglut2 and vgat are shown in Supplementary Table 1.2.

Peripheral blood mononuclear cells (PBMC) were isolated from venous blood of SLE patients or healthy controls using Ficoll-paque density gradient centrifugation. Purified CD4⁺ T cells were negatively isolated from PBMCs by CD4⁺ T-cell isolation kits (STEMCELL Technologies, Vancouver, Canada) according to the manufacturer’s protocol. CD4⁺ T cell purity was routinely > 90% as verified through flow cytometry. The cells were then cultured in Xvivo 15 medium (Lonza, Walkersville, MD, USA) supplemented with 10% human AB serum (Valley Biomedical, Winchester, VA, USA) at 37 °C with 5% CO₂. The treatments of the cells were: TNF-α (HY-P7058, MedChemExpress, NJ, USA), 10 ng/ml, 24 h; IL-6 (HY-P7044, MedChemExpress), 10 ng/ml, 24 h; 17β-estradiol (estradiol/E2) (HY-B0141, MedChemExpress), 100 nmol/L, 24 h; Lipopolysaccharides (LPS) (L8880, Solarbio, Beijing, China), 100 ng/ml, 24 h; ultraviolet B (UVB), 50 mJ/cm2 [10 (link)]; hydroxychloroquine sulfate (HCQ sulfate) (HY-B1370, MedChemExpress), 6 μg/ml, 24 h; 5-Azacytidine (5-aza C) (HY-10586, MedChemExpress), 1 mM, 24 h; prednisolone (HY-17463, MedChemExpress), 10 ng/ml, 24 h; AZD9496 (HY-12870, MedChemExpress), 5 nM, 24 h.