Figure 7

p45 NF-E2 regulates syncytiotrophoblast differentiation by SENP1-mediated GCM1 desumoylation in human trophoblast cells. (a–d) Decreased GCM1 sumoylation and enhanced GCM1–SENP1 interaction in 8-Br-cAMP-treated BeWo cells and primary human villous trophoblast cells (pHVT; a and b) and p45 NF-E2-deficient (c and d) BeWo cells; immunoprecipitation using the anti-GCM1 antibody followed by immunoblotting ((a and c) representative immunoblots; (b andd) bar graph summarizing results). (e and f) Overexpression of p45 NF-E2 induces GCM1 sumoylation while reducing the GCM1–SENP1 interaction both in the absence or presence of 8-Br-cAMP; immunoprecipitation using the anti-GCM1 antibody followed by immunoblotting ((e) representative immunoblots; (f) bar graph summarizing results). (g and h) Reduced GCM1 sumoylation and enhanced SENP1-GCM1 interaction in placenta tissue lysates obtained from women with pregnancies complicated with IUGR compared with controls, immunoprecipitation using anti-GCM1 antibody followed by immunoblotting ((d) representative immunoblots; (e) box-plot summarizing results). Mean±S.E.M. (b, d,f and h); results of at least five independent repeat experiments shown (b, d and f); *P<0.05 (t-test in b,d and h; ANOVA in f)