GSM9420676: CTL_1; Mus musculus; RNA-Seq
For single-nucleus sequencing, placentas from saline-exposed and poly(I:C)mod-treated male mice (n = 3) were snap-frozen in liquid nitrogen. Frozen tissues were thawed in 1 ml of high sucrose buffer (0.25 M sucrose, 25 mM KCl, 5 mM MgCl₂, 20 mM Tricine-KOH, pH 7.8, 0.15 mM spermine tetrahydrochloride, 0.5 mM spermidine trihydrochloride, 1 mM DTT) in presence of RNAse inhibitors. The samples were mechanically dissociated using a dounce homogenizer containing IGEPAL CA-630 (Sigma-Aldrich). Nuclei were collected from interface of 30% and 40% of iodixanol after 18 minutes of density gradient ultracentrifugation at 10,000 g. Purified nuclei were counted using a hemocytometer. Nuclei were extracted from placenta samples and processed using the 10x Genomics Chromium System. Libraries were sequenced using the Nextseq platform at the Centre for Applied Genomics (TCAG; The Hospital for Sick Children) in Toronto, ON, Canada.