Extended Data Fig. 2: Hydroxyestrogens are anti-inflammatory in vivo.

a. Representative gating strategy for identifying forward scatter/side scatter (FS/SS) live gate+, DAPI-, CD45+, F4/80+CD11b+ visceral white adipose tissue (vWAT) macrophages for sorting and flow cytometry analysis. b. vWAT mass in mice after 30 days HFD feeding and EtOH control or estrogen injections. n = 5 mice per group. c. vWAT macrophage cellularity in mice after 30 days HFD feeding and EtOH control or estrogen injections. n = 5 mice per group. d. Relative expression* of select pro-inflammatory genes in vWAT macrophages from HFD-fed mice injected with EtOH, 4-OHE1, or E2 (*log2-transformed RPKM centered on the mean of each gene). n = 5 mice per group. e. Blood glucose levels at 30min and 1h post-glucose injection in mice from Fig. 2f. n = 10, 10, and 11 mice per condition. f. qPCR for Nos2 expression in splenocytes isolated from mice in Fig. 2h. n = 3, 5, 5, and 5 mice per condition. For bar graphs, each data point is an independent biological replicate, and data is represented as mean ± SEM. All P values from unpaired, two-sided Student’s T Test (planned comparisons). HFD chronic inflammation model was performed once for transcriptional profiling (a-d), and a second time for metabolic studies (e,f).