Fig. 3: miR-200b binds to 3ʹ-UTR of Ambra1 mRNA and inhibits Ambra1 translation.

A Schematic representation of Ambra1 mRNA, depicting a potential miR-200b binding site (BS) located in Ambra1 3′-UTR. B Western blot of Ambra1 in E9.5 mouse embryos and quantification of Ambra1 protein in the control and NTD groups (n = 9). C Abundance of miR-200b and U6 examined by RT-qPCR after 48 h of biotin-miR-200b transfection. D Ambra1 mRNA levels detected in the lysates precipitated by biotin-miR-200b and the total input. E Schematic of different Ambra1 luciferase reporter plasmids. Luc Luciferase, BS miR-200b-binding site, WT wild-type, Mut mutation. F Relative luciferase activity levels of 3ʹ-UTR-Luc, WT-Luc, and Mut-Luc reporters after miR-200b transfection. Levels of miR-200b (G) and Ambra1 protein (H) in C17.2 cells transfected with the indicated dose of the miR-200b mimic or the control mimic for 48 h. Changes in miR-200b (I) and Ambra1 protein (J) expression after transfection of different doses of the miR-200b inhibitor or the control inhibitor. *P < 0.05 vs. control group.