GSM9150539: Long ascending propriospinal neurons 1; Mus musculus; RNA-Seq
Mice that received lumbar or cervical bilateral injections of Rabies-H2BCherry were sacrificed by decapitation. The cervical (C1 to T1) or lumbar segments (L1 to L6) of the spinal cord were cut into small pieces and placed in a Dounce homogenizer filled with ice-cold homogenization buffer. The tissue was manually homogenized with five strokes of the loose pestle, followed by 10–15 strokes of the tight pestle. Subsequently, the solution containing the dissociated nuclei was filtered through a 40 μm filter into a sorting tube and DAPI was added to a final concentration of 1 μM to label the nuclear population. Nuclear mCherry+/DAPI+ neurons were sorted into 96-well plates using BD FACSAria Fusion Single-nucleus RNA libraries were prepared following the CEL-Seq2 protocol (https://doi.org/10.1186/s13059-016-0938-8.)