Figure 6.

The HOTAIR/PI3K-AKT/MMP2 Axis Is Functionally Important for Regulating Trophoblast Invasion

(A and B) ELISA analysis of MMP2 expression in HTR-8 cells which were transfected with siCtrl or siHOTAIR (A), and vector or HOTAIR-overexpressing vector (B) for 48 hr. (C) Serum-free culture medium of HTR-8 cells transfected as indicated with siCtrl, siHOTAIR, and vector or HOTAIR overexpression plasmid was collected for gelatin zymography assay. (D) Extravillous explants from healthy controls (6–10 weeks) were maintained in culture on Matrigel. Serial pictures of the explants incubated with DMSO or LY294002 were taken under a light microscope after 24 and 72 hr of culture (original magnification ×100). (E and F) The supernatants were collected for gelatin zymography assay and ELISA. (G) Extravillous explants were cultured on Matrigel for 72 hr. Immunofluorescence staining using anti-MMP2 antibodies showed an obvious decrease in MMP2 protein levels in the LY294002-treated group compared to the DMSO-treated group. Green fluorescence signals indicate bound anti-MMP2 antibodies, red indicates CK7 staining, and the DAPI-stained nuclei are blue. Scale bars, 25 μm.