CLOCK Promotes Endothelial Damage by Inducing Autophagy through Reactive Oxygen Species

Figure 4

3-MA abolished the effects of CLOCK under hypoxia condition. CLOCK overexpressed HUVECs were pretreated with 3-MA (5 nM) for 2 h, followed by hypoxia treatment for 48 h. (a) Quantification of the number of autophagosomes. (b) The punctate GFP-LC3 dots were measured. (c) CCK-8 assay was used to assess the cell vitality. (d) Cell apoptosis was determined by a flow cytometry assay. (e) Cell migration was examined using transwell assays. (f) Bar graph showed the relative ROS level. (g) Immunoblotting for CLOCK, LC3, and Beclin1 expression in HUVECs as indicated treatment. , .

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