Fig. 1: Strategy for single nucleus RNA sequencing of spinal cord cholinergic neurons. | Nature Communications

Fig. 1: Strategy for single nucleus RNA sequencing of spinal cord cholinergic neurons.

From: Single nucleus RNA-sequencing defines unexpected diversity of cholinergic neuron types in the adult mouse spinal cord

Fig. 1

Spinal cords were extruded from Chat-IRES-Cre::CAG-Sun1-sfGFP mice and separated into 3 regions (Cervical, Thoracic, and Lumbar/Sacral) that were processed separately (1). Image from a whole cleared spinal cord (SC) immunolabeled for GFP (Supplementary Movie 1). Tissue was lysed (2) and nuclei were isolated by density gradient centrifugation (3). Nuclear suspensions (4) were sorted using fluorescent-activated cell sorting (FACS) (5) to select singlet GFP-positive DRAQ5-positive nuclei (6) that were processed for single nucleus RNA sequencing using the 10X Genomics Chromium platform (7). Single nucleus cDNA libraries were sequenced together on an Illumina HiSeq (7), then analyzed (8). Figure created with BioRender.com. Scale bars, (1) 200 µm, (5) 50 µm.

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