Endosome-associated lc3b mRNA is translated in axons and the protein contributes to endosomal clearance

The primary objective of this project was to identify transcripts in close proximity to endosomes using the APEX-seq approach. To achieve this, we employed SH-SY5Y cells expressing APEX2-NES, APEX2-RAB5A, or 2xFYVE-APEX2 constructs. Biotinylation of RNAs was validated by dot blot, and biotinylated RNAs were enriched via streptavidin immunoprecipitation and subsequently analyzed by RNA sequencing. This approach enabled us to identify mRNA encoding the LC3B protein in association with endosomes.