Figure 8.

Blocking of actin polymerization inhibits while calcium and ischemia promotes trophoblastic PP13 release. (A) PP13 content of BeWo cell culture supernatants of non-transfected controls, as well as LGALS13-transfected, untreated or Latrunculin B-treated cells were measured by ELISA (left). LGALS13-transfected, untreated or Latrunculin B-treated cells were stained with anti-PP13 (green) and nuclei were counterstained with DRAQ5 (blue) followed by confocal microscopic analysis (right; 40x magnifications). The disruption of the actin cytoskeleton with Latrunculin B treatment decreased PP13 release from BeWo cells. (B) LGALS13-transfected BeWo cells were treated with calcium ionophore to increase intracellular calcium level, or kept under ischemic stress to mimic placental milieu in preterm preeclampsia. (Left) PP13 content of BeWo cells and cell culture supernatants of non-transfected controls, as well as LGALS13-transfected, untreated, calcium ionophore-treated or ischemia exposed cells were measured by ELISA. (Left) Representative confocal images of LGALS13-transfected control, calcium ionophore-treated or ischemia exposed BeWo cells immunostained with monoclonal anti-PP13 antibody (green) and counterstained with DRAQ5 (red) nuclei dye. Either ionophore treatment or ischemia induced the release of PP13 from BeWo cells. Figures were published in Ref. (73). Kind permission for the reuse and modification of the figures was obtained from Elsevier.