Journal: bioRxiv
Article Title: Disrupted O-GalNAc glycosylation as a mechanism and biomarker of SLC35A2 -associated epilepsy
doi: 10.64898/2026.03.02.708854
Figure Lengend Snippet: A) Surgically resected tissue from a SLC35A2 -associated focal epilepsy patient (VAF = 3.4-5.2%) were analyzed for VVA positivity via lectin-based histochemistry. Using laser capture microdissection (LCM), VVA+ areas were isolated from VVA-areas and both were re-sequenced for the patient specific variant. B) Variant allele frequency (VAF) from areas positive and negative for VVA binding was measured using amplicon sequencing of DNA isolated via laser capture microdissection (LCM). Areas of diffuse VVA+ show enrichment for the patient-specific variant in SLC35A2 , which were absent in both VVA-areas and reference. C) Image of the SLC35A2- associated epilepsy tissue sample stained with VVA, visualized using DAB histochemistry and counterstained with hematoxylin/eosin. Scale bar = 2000 μm. D) High magnification images from (C) of areas of both low density VVA+ with individual cells (i), diffuse high density VVA+ (ii), and hypercellularity in white matter (iii). Scale bar = 100 μm.
Article Snippet: Lectin binding was visualized by incubating sections for two minutes in complete ImmPACT DAB reagent (VectorLabs, #SK-4105) and immediately rinsing for 5 minutes with two changes of tap water.
Techniques: Laser Capture Microdissection, Isolation, Variant Assay, Binding Assay, Amplification, Sequencing, Staining