SNAT - RNA sequencing of sorted Schwann cells as a single mixed population, enriched in myelinating Schwann cells, or enriched in not-myelinating Schwann cells

RNA sequencing of sorted mouse Schwann cells expressing YFP due to P0Cre-mediated recombination. The plotting of side scatter versus eYFP fluorescence generates 2 semi-detached eYFP+ clouds of cells, which further enables the enriched selection of myelinating Schwann cells, and of not-myelinating Schwann cells. Schwann cells were acutely extracted from P5 mouse sciatic nerves. These Schwann cells, besides of beta-actin dsred, also express eYFP upon P0cre-mediated recombination. Following extraction, Schwann cells were sorted via FACS. Plotting side-scatter versus eYFP fluorescence allows the selection of eYFP positive Schwann cells (as a mixture of myelinating and not-myelinating cells) from eYFP negative debris and non-Schwann cells in the nerve. Furthermore, 2 semi-detached eYFP+ clouds of cells could be detected at this mouse age, which further enabled the selection of populations either enriched in myelinating Schwann cells, or enriched in not-myelinating Schwann cells. For each gating strategy (mixed, myelinating, and not-myelinating), 4 independent biological samples were sequenced, 2 derived from male mice and 2 derived from female mice.