Figure 4.

ANGPTL4 is essential for rosiglitazone-induced trophoblast cell survival and proliferation. (a–c) HTR8/SVneo cells transfected with control siRNA (si-Con) or ANGPTL4 siRNA (si-ANGPTL4) were stimulated with 1 μM rosiglitazone in the presence of 150 μM hydrogen peroxide. Control cells were treated with 1 μM rosiglitazone or 100 nM recombinant human ANGPTL4 (rhANGPTL4) in the presence of 150 μM hydrogen peroxide. A TUNEL assay was performed to evaluate the rate of cellular apoptosis. Caspase-3 expression was detected by qRT-PCR, and the expression of caspase-9, cleaved caspase-9, caspase-3, cleaved caspase-3, cleaved PARP and Bax was measured via western blot analysis. The data are shown as the means±S.E.M. **P<0.01, ***P<0.001 compared with control or si-Con. (d–f) HTR8/SVneo cells transfected with si-Con or si-ANGPTL4 were stimulated with 1 μM rosiglitazone. Control cells were treated with 1 μM rosiglitazone or 100 nM rhANGPTL4. A CCK-8 assay was performed to examine cell proliferation. Cyclin D1 mRNA was assessed by qRT-PCR, and the expression of Bcl-2, pHH3, Cyclin D1 and c-Myc was determined by western blot analysis. The data are shown as the means±S.E.M. *P<0.05, **P<0.01 and ***P<0.001 against control or si-Con. (g) The expression of CK7, PPARγ, ANGPTL4, Cyclin D1 and caspase-3 in placental tissues was further analysed by IHC in PE subjects (n=30) and normal controls (n=30). Representative images were captured at × 200 magnification