In vitro decidualized hESCs ECM modulators expression and Chip experiments at the TIMP-1 and TIMP-3 gene promoters.

<p><b>A</b>, Quantitative PCRs for MMP-2, MMP-9, TIMP-1, TIMP-3, uPA and RECK expression levels in 5 day-cAMP+MPA-treated hESCs.. <b>B</b>, Extracellular TIMP-1 and TIMP-3 Western blot analyses of the protein extracts of 24 hour-conditioned media from hESCs treated with cAMP and MPA for 5 days. Ponceau staining of the membranes was used as a protein loading control. Only the 60 kDa band of the ponceau gel is shown. Representative image of three different analyses is shown. <b>C</b> and <b>D</b>, Chromatin immunoprecipitation experiments using antibodies against global H3 and H4 acetylation were carried out on the TIMP-1 and TIMP-3 genes in 24 hour-cAMP+MPA-treated hESCs. Relative amounts of immunoprecipitated TIMP-1 and TIMP-3 promoter sequences compared to input chromatin were determined by real-time PCR. <b>E</b>, Quantitative PCR for the TIMP-1 and TIMP-3 expression levels in 48 hour-cAMP+MPA, 1 µM TSA and cAMP+MPA+1 µM TSA-treated hESCs. Note that there is no significant difference between these two conditions at the level of the TIMP-1 and TIMP-3 gene expressions. Data represent the mean of three independent experiments. Statistical analysis,* p<0.05, ** p<0.01, a p<0.05, b p>0.05.