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When segmentation was incomplete due to weak fluorescence signals, missing nuclei were manually added

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To investigate whether this reduction in membrane-bound -SNAP occurs in a particular membrane subtype (i.e., plasma membrane or intracellular membranes), we subjected homogenates from E14.5 WT and hyh brains to a subcellular fractionation protocol that allows fractionating cytosol from intracellular (light) membranes (LM) and plasma membranes (PM)enriched fractions (Supplementary Fig