Subcellular localization of GFP-Us9 and phospho-GFP-Us9 in axons.
A.) Representative IF visualization of total GFP-Us9 in axonal vesicles and phosphorylated Us9 subspecies detected by phospho-specific mouse monoclonal 2D5E6 at 8 hours post-infection with PRV 340. Discrete puncta are highlighted: arrows indicate dual positive GFP-Us9 (+)/Alexafluor 568 (+) structures, triangles denote single positive GFP-Us9 (+) structures. B.) Quantitation of phosphorylation status of Us9 puncta in axons. A total of 626 GFP-Us9 puncta were scored in axons and the percentage phosphorylated determined through IF. GFP-Us9 puncta were counted across 3 biological replicates.