Fig. 2.

Keap1-Nrf2 PPI inhibitors downregulate estrogen responsive genes in E2-stimulated MCF-7 cells. A. The mRNA expression of estrogen responsive genes at 16 hours after treatment with 10 μM LH1092 or 100 nM CDDO-IM with or without 100 pM E2. B. The mRNA expression of estrogen responsive genes at 48 hours after treatment with 100 pM E2 in the presence or absence of 10 μM test compounds (LH1092, LH1093, LH1095 and LH1101) or 100 nM CDDO-IM. The mRNA levels were analyzed by RT-qPCR. GAPDH was used as an internal control for gene expression. The data are represented as mean ± SEM of mRNA fold changes (n = 6). **p < 0.05, significant difference compared to DMSO control. *p < 0.05, significant difference compared to E2 control. C. MCF-7 cells were treated with 10 μM LH1092, 10 μM LH1095 and 100 nM CDDO-IM in the presence of 100 pM E2 for 24 hours. The protein levels of PGR, TFF1/pS2, CTSD, and c-MYC were determined by Western blot analysis. β-actin was used as a loading control. Quantification of protein levels from the Western blots were analyzed using ImageJ program.