Fig 1: Characteristics of cultured human monocyte-derived macrophages. (A) Example of a monolayer of human monocyte-derived macrophages (hMDM) grown on ultralow adhesion plastic. (B) Basal hMDM secretome from 5 independent mixed sex cultures detected by RayBiotech human antibody array L-507 illustrating a mixed inflammatory and anti-inflammatory cytokine profile. (C) hMDM cell lysates analyzed by ELISA express both inflammatory (TNF-a, IL-6, IL-1ß, MCP-1, IFN-?) and anti-inflammatory (IL-13, TGF-ß1, IL-1ra) mediators with no evidence of sex differences at baseline. Each hMDM culture is shown as a single symbol (n = 18), obtained from at least 3 male or 3 female donors. (D) hMDM surface markers determined by flow cytometry using direct immunofluorescence in 4-7 independent mixed sex cultures. (E) Western blot image shows that hMDM from both male and female donors (n=9 cultures from 3 donors/sex) lack ?-aminobutyric acid type A (GABAA) receptor subunits a1, a2, a4, and d at baseline (CTL) or after treatments with lipopolysaccharide (LPS) and/or allopregnanolone (3a,5a-THP). As a positive control, the amygdala from male and female alcohol preferring P rats intraperitoneally injected with vehicle (45% w/v 2-hydroxypropyl-ß-cyclodextrin; 30 min) or allopregnanolone (15 mg/kg; 30 min) was used. (F) Chemical structures of endogenous neurosteroids allopregnanolone and 3a,5a-THDOC (tetrahydrodeoxycorticosterone) and the synthetic 1,2,5-triazole analog of the allopregnanolone, SGE-516. 3a,5a-THDOC differs from the allopregnanolone by a C-21-hydroxyl group at the D-ring. SGE-516 differs from the allopregnanolone by a C-3 cis-methyl and cis-hydrogens at C-5 and C-19, and a C-21-1,2,5-triazole group at the D-ring.