GSM8978429: LVCP, postnatal day 11 RNA, n=2; Mus musculus; RNA-Seq
Single-cell RNA sequencing was performed on lateral ventricle choroid plexus (CP) samples from mice at postnatal days P2-3, P11, and young adults (10 weeks), while single-cell multiome included P11 and young adult mice. CPs were isolated from perfused mouse brains, pooled (3–4 CPs/sample from both sexes), and enzymatically and mechanically dissociated through a multi-step protocol with pronase (0.1 mg/ml)  for 15min, Enzyme P mix (adult brain dissociation kit, 130-107-677, Miltenyi Biotec) for 30 min, and trypsin (0.025%) for 15min, with mechanic digestion every 5min. Cells pellets were resuspended in PBS-2%BSA. The Chromium Next GEM Single Cell 3ʹ v3.1 kit (10x Genomics) was used for scRNA-seq, following the manufacturer's guidelines. Single-cell multiome required additional steps of nuclei extraction using lysis buffer (IGEPAL (CA-630), 0.01% Tween-20, 0.001% Digitonin, 1% BSA, DTT 200mM, 10 mM Tris-HCl pH 7.4, 10 mM NaCl, 3 mM MgCl2 and RNAse inhibitor). After washing and centrifugation, an additional lysis step with Nuclei EZ buffer was performed (Sigma, #N3408). Nuclei were washed again and resuspended in diluted nuclei buffer (10x Genomics). A portion was used for viability counting, and the rest was prepared for tagmentation. The 10x Genomics Chromium Next GEM Single Cell Multiome ATAC + Gene Expression kit was used, following the manufacturer's protocol. 10x protocol to snMultiome