Extended Data Fig. 2: Neuronal nuclei FACS gating strategy.
From: Neuronal somatic mutations are increased in multiple sclerosis lesions

The presented gating strategy pertains to chronic lesion tissue from MS10, a PPMS case with excess sSNVs in lesion vs. NA tissue (Table 1, Fig. 2c). (a) A proportion of chilled pre-filtered nuclear preparation was stained with Hoechst 33342 and run through the FACS machine to identify DNA-containing nuclei. (b) NeuN stained nuclei mapping to the same position were gated and their size measured using forward scatter (FSC-A) and side scatter (SSC-A). (c) Singlet nuclei were identified and were gated further to remove auto-fluorescent (AF) particles (d). (e) Resulting neuronal nuclei were separated further based on strength of NeuN staining. (f) Single neuronal nuclei from the highest staining proportion, NeuN + +(hi), were sorted for whole genome amplification and snWGS.