Fig. 3

Effects on ER target genes expression and uterine tissue of SCR-6852. a Differential expression analysis from RNA-seq. MCF7 with hormone-deprived pre-treatment were administrated ER ligands or not present with 1 nM E2 stimulation for 24 h incubation in a 12-well plate. Total mRNA was isolated from treated cells and sequenced on an Illumina HiSeq X Ten platform. FPKM of each gene was calculated using Cufflinks and Differential expression genes analysis by using the DESeq (2012) R package. Four separate clusters were obtained. Rows show genes up- or downregulated by E2 relative to control (> twofold, FDR < 0.05), or by each ER ligand. b Summary of the transcriptional consequences. In orange is the percentage of upregulated genes compared with E2 in total test genes by experimental ligand (> twofold, FDR < 0.05), and those in blue are downregulated. c ER-target gene GREB1 or AGR3 expression analysis. T47D EFM-19 and HCC1500 cells were grown in RPMI media supplemented with 10% charcoal-stripped FBS for 14 days and then treated with ER antagonist (1 μM) or DMSO in the presence E2 (1nM) stimulation for 24 h before RNA isolation and gene expression analysis as described in methods. Asterisks show one-tail t-test p-values. P-values: * P < 0.05; ** P < 0.01. d-f Effect of SCR-6852 on uterine tissue. 21 days old SD rats were dosed with a vehicle, 60 mg/kg tamoxifen or 10 mg/kg SCR-6852 orally every day for three days. The uteruses were harvested 24 h after the final doses and stained by toluidine blue. Endometrial height was assessed from the basement membrane to the luminal border (scale bar in green lines: 200 μm), Arrows indicate the uterine epithelium (e). Wet uterine weight normalized to body weight and Endometrial thickness were digitally measured and plotted in the graph in figure (d and e). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001