Fig. 5.

Estrogen supplementation induces autophagy via estrogen receptor α. (A) Representative western blots and relative quantitative analysis of ERα and ERβ in cardiac aorta of three groups of mice. (B) Representative western blots and relative quantitative analysis of ERα and ERβ in HVUECs cells. (C) Representative western blots and relative quantitative analysis of ERα and ERβ with or without 12 h of application of 60 nM Hcy to HUVECs in the presence or absence of 10 nM E₂ or/and 10 μM MPP. (D) Representative western blots and relative quantitative analysis of ERα and ERβ with or without 12 h of application of 60 nM Hcy to HUVECs in the presence or absence of 10 nM E₂ or/and 10 μM PHTPP. (E) After transfection of HUVECs with GFP-LC3 adenovirus, immunofluorescence images of cells were taken with or without 60 nM Hcy for 12 h and in the presence or absence of 10 nM E₂ or/and 10 μM MPP. DAPI was used to stain the nucleus. Mean optical density analysis of GFP was performed. Scale bar: 50 μm. (F) After transfection of HUVECs with GFP-LC3 adenovirus, immunofluorescence images of cells were taken with or without 60 nM Hcy for 12 h and in the presence or absence of 10 nM E₂ or/and 10 μM PHTPP. DAPI was used to stain the nucleus. Mean optical density analysis of GFP was performed. Scale bar: 50 μm. In all experiments, n = 6. **P < 0.01.