A gene network of uterine luminal epithelium organizes mouse blastocyst implantation

RNA-seq analyses using embryos and uterine luminal layers collected from female mice on day 3 am, day 4 am, day 4 pm and day 5 am of pregnancy. Wildtype C57BL/6N females were used. The day when the vaginal plug was detected was considered day 1 of pregnancy. Pregnant mice were euthanized by cervical dislocation on designated days of pregnancy for the evaluation of pregnancy phenotypes and sample collection. On day 3, oviducts were flushed to confirm presence of 8 cells. On day 4, one uterine horn was flushed with saline to confirm the presence of blastocysts and the other side of the uterine horn was snap-frozen for cryosectioning. On day 5, mice were treated with 1% Chicago blue in saline i.v. to visualize implantation sites. Laser microdissection was performed to collect luminal layers.