Figure 3. TP53BP2 enhances trophoblast autophagy by regulating Beclin-1 expression. (A) Volcano plot of 2081 DEGs in HTR8/SVneo cells transfected with sh-TP53BP2. The red dots and green dots indicate upregulated and downregulated gene expression, respectively (|fold change|≥2.0, P ≤0.05), and the black dots indicate unchanged genes. (B) Heatmap of the top 10 downregulated autophagy-related genes in placentas from preeclamptic rats injected with AAV-shTP53BP2 . (C) qRT‒PCR validation of the top 10 downregulated genes. (D) The expression of Beclin-1, LC3B-II, p62 and TP53BP2 in HTR8/SVneo and JEG-3 cells transfected with Ad-TP53BP2 and/or sh-Beclin-1 under hypoxia was detected by western blotting. (E) Coimmunoprecipitation (co-IP) assay followed by immunoblotting showing the interactions of Bcl-2 with TP53BP2 and Beclin-1 in HTR8/SVneo and JEG-3 cells under hypoxia. The cell lysates were subjected to immunoprecipitation with an anti-Bcl-2 antibody.(F) Co-IP assay followed by immunoblotting showing the interactions of Bcl-2 with TP53BP2 and Beclin-1 in HTR8/SVneo and JEG-3 cells transfected with sh-TP53BP2 under hypoxia. The data are presented as the means ± SDs. *P <0.05,**P <0.01,***P <0.001;#P <0.05,##P <0.01.