Establishment of pleckstrin homology-like domain, family A, member 2 (PHLDA2) overexpressing cell lines; PHLDA2 overexpression inhibits cell proliferation. (A) JEG-3 cells and primary trophoblasts were infected with lentivirus overexpressing PHLDA2 or containing control vector. Total proteins were extracted after 48 h of infection and the concentrations were measured. Subsequently, western blot analysis was performed to detect the expression of PHLDA2 in JEG-3 cells and primary trophoblasts. PHLDA2 protein expression was normalized to β-actin expression. (B) Total RNA was isolated from the cells in each group after 48 h of infection. RT-qPCR was performed to detect PHLDA2 mRNA expression in JEG-3 cells and primary trophoblasts. (C) JEG-3 cells and primary trophoblasts were seeded in a 96-well plate. The effects of PHLDA2 on cell proliferation ability were analyzed by CCK-8 assay at 0, 24, 48 and 72 h. The results are expressed as the means ± SD. *P<0.05 and **P<0.01, compared with the control vector groups.