Extended Data Fig. 3: Purity assessment of neuronal nuclei.
From: Neuronal somatic mutations are increased in multiple sclerosis lesions
(a) Droplet Digital PCR (ddPCR) was performed on total RNA/cDNA from pooled FACS sorted NeuN-positive (+) nuclei from MS chronic (CL) lesion tissue and matched normal appearing (NA) tissue from a single MS case and NeuNhi (high-expressing, ++) nuclei from cerebellum (Cb) tissue from a single control (b) FACS gating strategy is shown for NeuN/H33342 stained nuclei preparations (y-axis) from MS6 chronic lesion (CL), matched MS6 NA tissue and CT5 frontal lobe (FL) tissue. For each tissue sample, FACS gates are shown for NeuNhi and NeuNlow nuclei populations, which were sorted into six pools for RNA extraction, cDNA synthesis and ddPCR (c) ddPCR results for the astrocyte marker, ALDH1L1, showing elevated expression in pooled MS6 CL NeuNlow nuclei and low, baseline expression levels in other nuclei pools.
