*4.2. Quantitative Reverse Transcription-PCR*

The myometrium was removed from implantation sites (IS) and total RNA was extracted from individual control and *Jag1*∆*EC* mutant IS using TRIzol (Invitrogen, Carlbad, CA, USA). Total RNA was treated with DNAse I (Invitrogen), reverse transcribed using qScript cDNA Supermix (Quanta Biosciences, VWR, Radnor, PA, USA) and gene expression were determined by quantitative (q) RT-PCR using the QuantiNova SYBR Green PCR Kit (Qiagen, Frederick, MD, USA) and the following primer sequences: *Jagged1* forward 50 -CTGCTTGAATGGGGGTCACT-30 , *Jagged1* reverse 5 0 -GCAGCTGTCAATCACTTCGC-30 ; *Dll4* forward 50 -GTTGCCCTTCAATTTCACCT-30 , *Dll4* reverse 5 0 -AGCCTTGGATGATGATTTGG-3; *Hey2* forward 50 -AAGCGCCCTTGTGAGGAAAC-30 *Hey2* reverse 5 0 -GGTAGTTGTCGGTGAATTGGAC-30 ; *Nrarp* forward 50 -GCG TGG TTA TGG GAG AAA GAT-30 , *Nrarp* reverse 50 -GGG AGA GGA AAA GAG GAA TGA-30 . Relative expression levels were quantified using the 2−∆∆Ct method and are expressed as fold change normalized to β-actin forward 5<sup>0</sup> -CGT GAA AAG ATG ACC CAG ATC-30 and β-actin reverse 50 -CAC AGC CTG GAT GGC TAC GT-30 .
