Transcriptional programs controlling closure of pallial germinal activity
Description
The dorsal subventricular zone (dSVZ) from P12 mice (2 males) of 2 independent experiments were dissected. During all stages of the dissociation protocol, the tissue was kept in aCSF solution containing 125 mM NaCl, 2.5 mM KCl, 1.25 mM NaH2PO4, 26 mM NaHCO3, 17 mM glucose, 1.25 mM CaCl2 and 1 MgCl2. The small dissected tissues were incubated in Papain during 20 minutes at 37°C. Following enzymatic digestion, cells were centrifuged for 5 minutes and the pellet was resuspended with DNAase/ovomucoid inhibitor according to manufacturer's protocol (Worthington). Cells were then centrifuged, triturated and resuspended on ice in Leibovit’z L-15 medium supplemented with 0.1% of bovine serum albumin. The cell suspension was finally filtered through a 70-μm cell strainer to remove aggregated cells. mRNA libraries were prepared according to 10x Genomics 3’ version 3 chemistry & Illumina protocols.