1. Briefly, CZI.files were converted into arivisSIS format with no compression, using Zen 3.2. The quantification pipeline in Arivis Vision4D 3.3.0 included: 1) a fluorescence threshold to exclude background (based on 2ry-only stained SCOs), 2) global enhancement;Simple Sharpening Filter
2. 88803) were added to the anti-SMN and anti-IgG reactions and incubated for 2 hours at 4?C with rotation. Following bead incubation, beads were separated from supernatant and washed 3x with pre-chilled wash buffer and 3x with pre-chilled 1x PBS for 10min each at 4?C with rotation. Following washes, proteins were eluted from beads with 2x Laemli Buffer (Bio-RAD, 1610737) with ?-mercaptoethanol and boiled at 95?C for 15minutes along with total lysate samples;Reactions were incubated overnight at 4?C with rotation, and then 15�L of Pierce Protein A/G Magnetic Beads