FIGURE 5.

Hypoxia induces Ser³²² phosphorylation in GCM1 in vivo. A, GSK-3β phosphorylates Ser³²² under hypoxia. 293T cells were transfected with pHA-GCM1 plus GL2 or GSK-3β siRNA. The transfected cells were subjected to hypoxic conditions for 24 h in the presence of MG132, followed by immunoprecipitation (IP) with HA mAb and immunoblotting (IB) with Ser(P)³²²-GCM1 Ab. B, Ser³²² phosphorylation in placental cells under hypoxia. BeWo31 cells were cultured under normoxia or hypoxia for 48 h and mock-treated or treated with the indicated combination of MG132 and LiCl, followed by immunoprecipitation with HA mAb and immunoblotting with HA, Ser(P)³²²-GCM1, or Ser(P)³²⁶-GCM1 antibody. C, GCM1 degradation under placental hypoxia contributes to the pathogenesis of preeclampsia.