Fig. 3

Role of ACNT4 in villous explants outgrowth. a IHC staining of ACTN4 in the human first trimester placenta villi. iEVTs were stained for HLA-G. Scale bars: 200 μm. b EdU labeling in explant cultures. Representative pictures showed EdU (pink) labeling in the CCTs of the anchoring villi after 72-h treatment of ACNT4 with siRNA. Nuclei were counterstained with DAPI (blue). Scale bars: 100 μm. c Outgrowth of induced EVTs in a villous explant culture model. Left panel, bright and fluorescent field views showing that small interfering RNA labeled with FAM successfully penetrated the villous explants. Right panel, the outgrowth of induced EVTs from villous explants treated with control siRNA (si-NC) or ACTN4 siRNA (si-ACTN4) for 24 and 48 h. Scale bar: 1000 μm. All data are presented as the means ± SEM of three independent experiments, ^*P < 0.05; ^**P < 0.01