Fig. 4: Role of ER in E₂β stimulation of CBS and CSE transcription.

(A) Human CBS and CSE promoter luciferase-reporter constructs were transfected into UASMCs. Twenty-four hours later cells were allowed to recover before treatment with vehicle or 10 nM E₂β with or without 1 μM ICI for 24 h. Promoter activity was determined. (B) UASMCs were treated with vehicle or 10 nM E₂β in the presence or absence of 1 μM ICI for 48 h. CBS and CSE mRNAs were determined by qPCR. (C) Cells were treated with vehicle, 10 nM E₂β, 10 nM agonist of ERα (PPT) or ERβ (DPN), or PPT+DPN to assess the role of ERα and ERβ agonists on CBS and CSE mRNA expressions. (D) Cells were treated with vehicle or 10nM E₂β in the presence or absence of 1 μM antagonist of ERα (MPP) or ERβ (PHTPP), or MPP+PHTPP to assess the role of ERα and ERβ antagonists on CBS and CSE mRNA expressions. Data (mean ± SEM) were collected from cells prepared from 3–5 different ewes. Bars with different letters differ significantly (P < 0.05), capital letters pertain to CBS and lowercase to CSE. *** P<0.001.