Fig 1.

GCM1 regulates placental cell invasion. (A) Expression of GCM1 in JAR and BeWo cells. JAR and BeWo cells were subjected to immunoblotting with GCM1 and β-actin Abs. (B) Stimulation of JAR cell invasion by GCM1. Mock (pCDH) and HA-GCM1-expressing (pCDH-HA-GCM1) JAR cells were plated in Matrigel-coated chambers for invasion analysis. Representative pictures of cells that migrated to and invaded the lower surface of filters are provided. Means and standard deviations (SD) obtained from three independent experiments are presented. (C) GCM1 knockdown suppresses BeWo cell invasion. BeWo cells stably expressing scrambled or GCM1 shRNA were subjected to invasion analysis as described for panel A. (D) Identification of GCM1 target genes. BeWo31 cells were treated with 50 μM forskolin for 24 h, followed by ChIP-chip analysis as described in Materials and Methods. HtrA4 was identified as a candidate GCM1 target gene with strong hybridization signals (red versus blue) in the region upstream of exon1 of the HtrA4 gene (right). PGF, encoded by a known GCM1 target gene, was also identified in the analysis (left).