Activin-Like Kinase 2 Functions in Peri-implantation Uterine Signaling in Mice and Humans
Figure 7
SMAD1,5,8 phosphorylation in hrBMP2-treated human endometrial stromal cells and SMAD1/5 and CEBPB ChIP experiments.
A) Phosphorylation of SMAD1,5,8 in hESC treated with 100 ng/ml hrBMP2 for 0 (non treated = NT), 30, 45, 60 minutes, 3 or 6 hours. B-ACTIN was used as loading control. B) Phosphorylation of SMAD1,5,8 in hESC transfected with NT (left) or ALK2 siRNA (right) and treated with 100 ng/ml hrBMP2 for 0 (NT), 30, 45 or 60 minutes. C) qPCR quantification (Sybr green) of DNA recovered after pull down with SMAD1/5 or CEBPB (E) antibody. We used a pair of primers (Table S1) specific for putative binding sites in proximity of the CEBPB (D) and PGR (F) genes. The quantification is given as fold enrichment of SMAD1/5- or CEBPB-pulled down vs. IgG-pulled down DNA. UTR = untranslated region used as negative control (primers targeting a fragment from a gene desert on chromosome 4).