Fig. 6

Quercetin inhibits AKT and ERK1/2 phosphorylation and signaling and promotes p53 (Ser46) phosphorylation. (A-B) Control endometrial stromal cells (eSCs) were treated with quercetin (Q, 25µM) for 4 h before analyzing cell lysates using a phospho-kinase array panel. Representative arrays (A) and quantification of array analytes/spots (B) are shown. Data for differentially expressed targets are presented as specific spot density normalized to the reference spot comparing Veh- vs. Q-treated (B). Each point represents the specific density of each duplicate analyte spot as a percentage of Veh-treated, with group median ± IQR shown for control- and endometriosis-eSCs. **p < 0.01. The remaining arrays are in Supplementary Fig. 7. (C-D) Control (CTRL) and endometriosis (ENDO) eSCs were treated with Veh or Q (25µM) for 4 h before western blotting for p-AKT, total AKT, p-ERK1/2, total ERK1/2, p-PRAS40, and total PRAS40. Representative blots and quantification of specific analytes are shown in (C) and (D), respectively. Band densities were normalized to GAPDH and shown as fold-change between Veh- vs. Q-treated eSCs in (D). Each point represents data from one individual’s eSCs, with median ± IQR shown for each group. *p < 0.05; ***p < 0.001; ****p < 0.0001. (E) Control-eSCs were treated with Veh or Q (25µM) for 0.5, 2, and 4 h before western blotting for phospho-AKT, total AKT, phospho-ERK1/2, total ERK1, phospho-PRAS40, and total PRAS40. (F-I) Control-eSCs were treated with vehicle (Veh), AKT inhibitor MK-2206 (MK, 1µM), quercetin (Q, 25µM) or Q + MK for 4 h prior to cAMP (F and H) or cAMP + MPA (G and I) stimulation; decidualization was assessed by measuring IGFBP1 (F and G) or PRL (H and I) by ELISA. Data are presented as fold-change in IGFBP1 or PRL over Veh-treated (Veh-treated = 1). Each symbol represents data from one individual’s eSCs, with median ± IQR shown for each group. *p < 0.05; **p < 0.01 vs. Veh-treated; ***p < 0.001 vs. Veh-treated; ns = non-significant vs. Q-treated